Cancer Research and Treatment

Original Article

Analysis of Cellular Changes Resulting from Forced Expression of Dickkopf-1 in Hepatocellular Carcinoma Cells: Mi Hee Kwack, Sun Young Hwang, In Seok Jang, Sang Uk Im, Jin Oh Kim, Moon Kyu Kim, Jung Chul Kim, Young Kwan Sung; Cancer Res Treat. 2007;39(1):30-36. Published online March 31, 2007; DOI: https://doi.org/10.4143/crt.2007.39.1.30

Purpose

Recent studies have shown that Dickkopf-1 (DKK-1) is overexpressed in some tumors, including hepatocellular carcinoma. However, the role of increased DKK-1 in these tumors is not known. In this study, the DKK-1 expression in hepatocellular carcinoma (HCC) cell lines was evaluated and the effect of DKK-1 overexpression in HCC cell lines was studied.

Materials and Methods

The expression of DKK-1 in hepatocellular carcinoma cell lines was evaluated by RT-PCR. Stable cell lines that overexpressed DKK-1 were established. Cell growth, adhesion, migration and invasion assays were performed.

Results

RT-PCR analysis showed that 5 out of 8 HCC cell lines expressed DKK-1. The forced expression of DKK-1 suppressed the growth of cells and increased the population of cells in the sub-G1 phase. In addition, DKK-1 reduced the cellular adhesion capacity to collagen type I and fibronectin, and it increased migratory capacity. However, overexpression of DKK-1 did not increase the invasion capacity of the HCC cell line.

Conclusion

Collectively, our data suggest that overexpression of DKK-1 affects the biology of HCC cells.

Citations

Citations to this article as recorded by

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Serum Dickkopf-1 as a Biomarker for the Diagnosis of Hepatocellular Carcinoma
Seung Up Kim, Jeon Han Park, Hyon-Suk Kim, Jae Myun Lee, Hyun Gyu Lee, Hyemi Kim, Sung Hoon Choi, Shinhwa Baek, Beom Kyung Kim, Jun Yong Park, Do Young Kim, Sang Hoon Ahn, Jong Doo Lee, Kwang-Hyub Han
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Serum level of DKK-1 and its prognostic potential in non–small cell lung cancer
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Review Article

Biology of SNU Cell Lines: Ja-Lok Ku, Jae-Gahb Park; Cancer Res Treat. 2005;37(1):1-19. Published online February 28, 2005; DOI: https://doi.org/10.4143/crt.2005.37.1.1

Abstract PDF PubReader ePub

SNU (Seoul National University) cell lines have been established from Korean cancer patients since 1982. Of these 109 cell lines have been characterized and reported, i.e., 17 colorectal carcinoma, 12 hepatocellular carcinoma, 11 gastric carcinoma, 12 uterine cervical carcinoma, 17 B-lymphoblastoid cell lines derived from cancer patients, 5 ovarian carcinoma, 3 malignant mixed Mllerian tumor, 6 laryngeal squamous cell carcinoma, 7 renal cell carcinoma, 9 brain tumor, 6 biliary tract, and 4 pancreatic carcinoma cell lines. These SNU cell lines have been distributed to biomedical researchers domestic and worldwide through the KCLB (Korean Cell Line Bank), and have proven to be of value in various scientific research fields. The characteristics of these cell lines have been reported in over 180 international journals by our laboratory and by many other researchers from 1987. In this paper, the cellular and molecular characteristics of SNU human cancer cell lines are summarized according to their genetic and epigenetic alterations and functional analysis.

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Original Articles

Detection of p16(INK4A) in the Mixed Cell Populations of Normal Peripheral Blood Mononuclear Cells and Cervical Cancer Cell Lines

Ji Young Kwon, Yoon Sung Jo, Ye Hoon Choi, Jong Gyu Chang, Ki Sung Ryu, Jong Gu Rha, Ku Taek Han

Cancer Res Treat. 2003;35(3):254-260.   Published online June 30, 2003

DOI: https://doi.org/10.4143/crt.2003.35.3.254

Abstract PDF

PURPOSE
Human papilloma viruses (HPVs) play a central role in the pathogenesis of neoplastic lesions of the uterine cervix. The viral oncoprotein HPV E6 degrades the p53 protein, and the HPV E7 protein inactivates pRB and increases the expression of the CDK inhibitor, p16(INK4A). We investigated the usefulness of p16(INK4A) as a biologic marker for the cervical dysplastic and neoplastic cells.
MATERIALS AND METHODS
We examined the expression of p16(INK4A) and cytokeratin in a mixed population of normal peripheral blood mononuclear cells (PBMC) and the cervical cancer cell lines (HeLa, SiHa, and CasKi) using flow cytometry. RESULTS: The DNA indices of the HeLa, SiHa and CasKi cell lines were 1.89, 1.53 and 1.75, respectively, indicating that these cells are aneuploid cells. Furthermore, the positive rate of p16(INK4A) expression was 86.7% for the HeLa mixed population, 85.6% for the SiHa mixed population, and 92.2% for the CasKi mixed population. According to the FL3A vs FL3W histogram, electrical gating of the HeLa, SiHa and CasKi mixed populations showed the expression levels of both cytokeratin and p16(INK4A) to be identical, at 86.6%, 84.8% and 85.0%, respectively. These findings revealed that almost all cells selected through electrical gating were cervical cancer cells originating from the epithelium and which expressed cytokeratin and p16(INK4A). On the other hand, when each mixed population was electrically gated for normal PBMC, we found that the PBMCs expressed neither cytokeratin nor p16(INK4A).
CONCLUSION
Using flow cytometry, we observed the enhanced expression of p16(INK4A) in cervical cancer cell lines. These RESULTS suggest the usefulness of p16(INK4A) for the selective detection of cervical dysplastic and cancer cells in the liquid-based samples, which are taken from the cervices and contaminated with blood and stromal cells.

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2-Methoxyestradiol: A Hormonal Metabolite Modulates Stimulated T-Cells Function and proliferation
J.G.Y. Luc, R. Paulin, J.Y. Zhao, D.H. Freed, E.D. Michelakis, J. Nagendran
Transplantation Proceedings.2015; 47(6): 2057.     CrossRef

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Retinoic Acid Enhances Drug-Induced Cell Death in Anticancer Drug-Resistant Cell Lines

Young Mi Whang, Yeul Hong Kim, Sang Won Shin, Byung Soo Kim, Jun Suk Kim, Young Do Yoo, Sun Hee Park

Cancer Res Treat. 2002;34(3):212-217.   Published online June 30, 2002

DOI: https://doi.org/10.4143/crt.2002.34.3.212

Abstract PDF

PURPOSE
Retinoids (RA), a group of vitamin A derivatives, is known to be important for regulation of normal cellular growth and differentiation. RA treatment of various cancers resulted in cell growth inhibition and apoptosis. Therefore, the chemotherapeutic and chemopreventative activities of various types of tumor have been examined. Biological actions of RA are mediated through nuclear receptors, including the retinoic acid receptors (RARs) and retinoid X receptors (RXRs). In this study, we examined the effect of all-trans-retinoic acid (atRA) as an anticancer drug-sensitiser in cancer cell lines and in its drug- resistant cancer cell lines MATERIALS AND METGODS: Cells were maintained by RPMI 1640 medium containing 10% fetal bovine serum. Cells were treated with 1 micro M atRA for 48 h, then with the desired concentration of anticancer drug for 24 h. Cell viability was measured spectrophotometrically at 540 nm using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. Western blot analyses were performed with the desired antibodies.
RESULTS
We investigated if pre-treatment with atRA enhanced the drug-sensitivity of various cancer cell lines to either 5-fluorouracil, adriamycin, or cisplatin. 5-FU (SNU638-F2) and CDDP-resistant cell (SNU638-Cis) lines, from a Korean gastric cancer cell line (SNU638) and the ADR-resistant cells (AD600) was established from a colon cancer cell line (SW620). Treatment of each cell line, with 1 micro M atRA, prior to drug exposure resulted in enhanced cell death in these cell lines. Furthermore, the effect of atRA on growth inhibition, in each drug-resistant cell line, was more obvious than in their parent cell lines. Increased activity of Transglutaminase II (TgaseII) and cleavage of Poly (ADP-ribose) polymerase (PARP) were also observed (western blot analysis CONCLUSION: Based on our data, we suggest that atRA enhances anticancer drug-induced cell death and reverses the drug-sensitivity of the drug-resistant cancer cell lines.

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All-trans Retinoic Acid Sensitizes Epithelial Ovarian Cancer to PARP Inhibition after Exposure to Cisplatin
Bingjie Mei, Junyang Li, Dengfeng Wang, Lu Feng, Jianming Huang, Guonan Zhang
Molecular Cancer Therapeutics.2025; 24(3): 453.     CrossRef

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5-FU Induces Apoptosis of Fas (+), HepG2 Cells Via Activation of Fas-mediated Caspase and Mitochondria Dysfunction

Channy Park, Kui hyun Yoon, Young Jin Lee, Yong Kweon Kim, Yee Cheon Choi, Jae Hoon Shin, Jeong Hwan Cho, RaeKil Park

Cancer Res Treat. 2002;34(2):128-138.   Published online April 30, 2002

DOI: https://doi.org/10.4143/crt.2002.34.2.128

Abstract PDF

PURPOSE
In order to investigate the role of Fas on the chemosensitivity of cancer cells in regards to chemotherapeutic agents, the Fas/FasL signaling pathway of apoptosis was explored in human hepatoma cells.
MATERIALS AND METHODS
Fas expression of hepatoma cells including Chang, Huh7, HepG2, and Hep3B cells, was determined by RT-PCR and flow cytometry analysis. Cell viability was measured by MTT assay and apoptosis was assessed by DNA fragmentation assay. The catalytic activity of the caspase-family proteases including caspase-3, 6, 8, and 9 proteases, was tested using fluorogenic biosubstrates. The expression of apoptotic mediators including cytochrome c, PARP, and Bcl2 family proteins were measured from cytosolic and mitochondrial compartments. Mitochondrial membrane potential was measured by fluorescence staining with JC-1, rhodamine 123.
RESULTS
Fas mRNA was constitutively expressed in Chang and HepG2 as defined as Fas (+) cells, but not in Huh7 and Hep3B cells, defined as Fas (-) cells. Fas (+) cells were markedly sensitive to 5-FU whereas Fas (-) cells were resistant and able to survive. 5-FU increased Fas expression of Fas (+) HepG2 cells and simultaneously resulted in apoptotic death, characterized by the ladder-pattern fragmentation of genomic DNA. Moreover, it increased the catalytic activity of caspase-8 protease, which eventually cleaved the Bid into truncated Bid which translocated into mitochondria only in Fas (+) cells. It also increased the caspase-9 protease activity with Bax expression, cytosolic release of cytochrome c, and mytochondrial dysfunction only in Fas (+) HepG2 cells. Furthermore, 5-FU increased the enzymatic activity of caspase-3 protease with PARP digestion in HepG2 cells.
CONCLUSION
5-FU exerted cytotoxicity against hepatoma cells via activation of Fas-mediated apoptotic signaling including caspase cascades and mytochondrial dysfunction. Our data suggests that Fas may be an important modulator of the chemosensitivity of cancer cells vis- -vis anticancer chemotherapeutic agents.

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Sex-dependent liver cancer xenograft models for predicting clinical data in the evaluation of anticancer drugs
Sungryong Oh, Joohee Jung
Laboratory Animal Research.2021;[Epub]     CrossRef
Lead bioactive compounds of Aloe vera as potential anticancer agent
Ranabir Majumder, Chandan Kanta Das, Mahitosh Mandal
Pharmacological Research.2019; 148: 104416.     CrossRef
Extract from Artemisia annua Linn? Induces Apoptosis through the Mitochondrial Signaling Pathway in HepG2 Cells
Bo Min Kim, Guen Tae Kim, Eun Ji Kim, Eun Gyeong Lim, Sang-Yong Kim, Young Min Kim
Journal of the Korean Society of Food Science and Nutrition.2016; 45(12): 1708.     CrossRef
Dexamethasone Inhibits TRAIL- and Anti-cancer Drugs-induced Cell Death in A549 Cells through Inducing NF-κB-independent cIAP2 Expression
Youn Seup Kim, Jae Seuk Park, Young Koo Jee, Kye Young Lee
Cancer Research and Treatment.2004; 36(5): 330.     CrossRef

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The Expression of Insulin-like Growth Factor (IGF), IGF- Binding Protein (IGFBP) and the Role of IGFBP-3 in the Korean Gastric Cancer Cell Lines

Dae Yeol Lee, Ho Keun Yi, Doo Hyun Yang, Pyung Han Hwang

J Korean Cancer Assoc. 2001;33(2):121-129.

Abstract PDF

PURPOSE
Insulin-like growth factor (IGF)-I and II are potent mitogens, postulated to exert autocrine and paracrine effects on growth regulation in human gastric cancer. In this study, we evaluated the expression of IGF-I, -II and IGFBPs in a panel of human gastric cancer cell lines. We also evaluated whether high expression of IGFBP-3 in human gastric cancer cells may increase the sensitivity to the anti-proliferative agents.
MATERIALS AND METHODS
10 human korean gastric cancer ceIl lines and 1 Caucacian gastric adenocarcinoma cell line were used for this study. IGF and IGFBP expressions were evaluated by RT-PCR. IGFBP proteins in conditioned media were detected by Western Ligand Blot. Cell survival after treatment of anti-proliferative agents was assessed by MTT assay.
RESULTS
IGF-I and II were expressed in all gastric cancer cell lines. In addition, IGF-I and II stimulated the proliferation of gastric cancer cells. The expression of IGFBP-2 was found in all gastric cancer cell lines. IGFBP-4 was expressed in the most of cell lines. IGFBP-3, -4 and -6 were expressed in about 50% of cell lines. The growth inhibition of IGFBP-3 expressing cells by anti- proliferative agents was more significant than that of IGFBP-3 nonexpressing cells. Cell growth inhibition with treatment of these agents was accompanied by increased IGFBP-3 mRNA level.
CONCLUSION
These data confirm that IGF-I, -II, and certain IGFBPs were expressed in gastric cancer cells, and gastric cancer cells show the differential growth inhibition by anti-proliferative agents. The differential growth inhibitory effect of anti-proliferative agents is, at least in part, mediated through up-regulation of IGFBP-3 expression.

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Growth Regulation of Ovarian Cancer Cells through the Inactivation of AP-1 by Retinoid Derivatives

Young Me Koh, Jong Sup Park, Sung Eun Namkoong, So Young Lee, Soo A Kim, Kyong Ja Hong, Soo Jong Um

J Korean Cancer Assoc. 2000;32(6):1043-1049.

Abstract PDF

PURPOSE
The growth regulatory effect of retinoid derivatives could be mediated by the transcriptional inactivation of AP-1 oncogenic transcription factor. By using ovarian cancer cell lines we were to investigate the cross-regulation mechanism between retinoids and AP-1.
MATERIALS AND METHODS
Cell proliferation assays were performed in 4 ovarian cancer cells (A2774, PA-1, OVCAR-3, SKOV-3) by increasing the concentrations of all-trans retinoic acid (ATRA), 9-cis retinoic acid (9RA), 13-cis RA (13RA), 4-hydroxyphenyl retinamide (4-HPR). Transient transfection and CAT ELISA were done to determine the selective activity of each retinoid on the RAR (alpha, beta, gamma), RXR (alpha, beta, gamma). and the negative activity on AP-1 (c-Jun).
RESULTS
Antiproliferative effect of 4-HPR (IC50; 0.7~2.7 micrometer) was more potent than those of other retinoid derivatives (IC50; 2.7~9.0 micrometer). To assess the anticancer mechanism, we examined the effect of 4-HPR on the transriptional activity of retinoic acid receptors (RAR/RXR) and of c-jun. Contrary to other retinoid derivatives that are active for RAR and RXR with some different levels, 4-HPR showed weak activity only for RARgamma. However, 4-HPR exerted the strongest suppression on AP-1 (c-Jun) activity.
CONCLUSION
Based on our results showing much 4-HPR's potent antiproliferative activity coupled with the most effectively inhibiting activity on AP-1 and minimum activity on RA receptor (selective for RARgamma) than other retinoid derivatives, we suggest that 4-HPR may be a novel, and very effective anticancer drugs for ovarian cancer.

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Phorbol Ester Induced - Apoptosis Mediated by Activating Serine Protease ( s ) and Caspase - 3 / CPP32 in SNU - 16 Human Gastric Cancer Cell Line

I C Park, M J Park, T B Choe, J J Jang, S I Hong, S H Lee

J Korean Cancer Assoc. 2000;32(2):270-278.

Abstract PDF

PURPOSE
Protein kinase C (PKC) is a family of phospholipid dependent serine/threonine protein kinases that have important role in differentiation, development and tumor promotion. PKC also has been reported to be implicated in the induction of apoptosis in a number of studies, but the efforts to define a role for PKC in the induction of apoptosis have been complicated by conflicting reports.
MATERIALS AND METHODS
To determine the effect of phorbol 12-myristate 13-acetate (PMA) on the induction of apoptosis, DNA fragmentation was detected by agarose gel electrophoresis and morphological changes of apoptotic cells were detected by Hoechst 33258 staining. For the detection of caspase-3/CPP32 activity, we used the enzyme substrate Ac-DEVD-pNa and anti- D4-GDI antibody.
RESULTS
In the present study, PMA, a PKC activator, induced apoptosis in SNU-16 human gastric cancer cell line, whose apoptosis was significantly inhibited by the PKC inhibitor, chelerythrine chloride. The caspase-3/CPP32 protease activity was increased in PMA-induced apoptosis. Furthermore, 4-(2-aminoethyl) benzenesulfonyl fluoride (AEBSF), a serine profease inhibitor, also significantly suppressed PMA-induced cell death in an upstream of caspase-3/CPP32.
CONCLUSION
These findings indicate that PMA induces apoptotic cell death in the SNU-16 adenocarcinoma cells through PKC activation, which activates AEBSF-sensitive serine proteases and caspase-3/CPP32. Therefore, our results suggest that PKC would be a potential target for induction of apoptosis.

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Chromosomal Aberrations in Ovarian Carcinoma Cell Line, SNU - S , Using Chromosome Painting

Jae Seong Kang, Dae Woon Kim, Yong Hyuck Chun, Sun Hwa Park

J Korean Cancer Assoc. 2000;32(1):120-128.

Abstract PDF

PURPOSE
The characterization of all recognizable chromosomal rearrangements was dis- turbed by technical limitation of conventional cytogenetic methods. Recently, the strong usefullness of generation of chromosome specific painting probes in identification of marker chromosomes has proven. This study was intended to analyze the chromosomal aberrations in human ovarian cancer cell line, SNU-8, by G-banding and multiple paintings.
MATERIALS AND METHODS
Human ovarian cancer cell line, SNU-8 was cultured and harvested for cytogenetic analysis. Routine karyotyping was performed. For complete analysis of chromosomal aberrations, human chromosome-specific painting probes were constructed from somatic hybrid cells. The origins of the unidentified marker chromosomes were analyzed by fluorescent in situ hybridization (FISH) with these painting probes.
RESULTS
All chromosome alterations were confirmed by the use of multiple chromosome paintings, which also demonstrated a number of additional alterations. SNU-8 had the karyotype 62-69,XXX, + der(1;10)(q10;p10),der(3;18) (q10;p10)X2,-4,+ 5,+ 7,del(9)(q21)X2,-11,-13,-15,-16,der(17;19)(q10;q10) X2, + 20,-22[cp51].
CONCLUSION
The chromosomal aberrations of SNU-8 cell line was effectively analyzed by FISH with these painting probes, and the approach methods of this study can be applied to cytogenetic analysis of chromosomal aberrations in the other cancers.

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Expressions of Cell Cycle Control Genes in Human Uterine Cervical Cancer Cells

Jung Geol Ahn, Tae Seong Lee, Jae We Cho, Won Ki Baek, Seong Il Suh, Min Ho Suh, Jong Wook Park, Soon Do Cha

J Korean Cancer Assoc. 2000;32(1):110-119.

Abstract PDF

PURPOSE
Recently, many aspects of biological functions of cyclins, cyclin-dependent kinases (CDKs), CDK inhibitors and Rb gene have been reported, and the cell cycle control genes are considered to act important roles in tumorigenesis. In this study, the expression patterns of major cell cycle control genes (cyclin A, B, C, Dl, E, E2F, p16INK4a, p21WAF1 and Rb) in various human cervical cancer cells were analysed to elucidate the impacts of the cell cycle control genes on the carcinogenesis of human cervical cancer.
MATERIALS AND METHODS
The expression patterns of major cell cycle control genes in HT-3, C33-A, HeLa, C4-II, SiHa and CaSki human uterine cervical cancer cells were analysed by using western blot and reverse transcription-polymerase chain reaction (RT-PCR).
RESULTS
In most of the cervical cancer cells tested, the overexpressions of cyclin A, E, E2F and markedly decreased expression of Rb tumor suppressor proteins were observed. By comparing RNA and protein expressions in each cancer cells, the mechanisms of increased expressions of cyclin A, E and decreased expression of Rb were elucidated as post-translational controls.
CONCLUSION
The cervical carcinogenesis caused by the altered expressions of the major cell cycle control genes can be hypothesized as follows: overexpressions of cyclin E and A cause acceleration of Rb phosphorylations and E2F overexpression; increased E2F function accelerates G1/S transition of the cells; compensatory increase of p16 expression cannot stop the cells in Gl phase because Rb expression is severely decreased; consequently, loss of Rb function, 61 shortening, inappropriate cell division and decreased function of the maintenance of genomic stability occur. In addition to these alterations, loss of p53 functions further accelerate instability of genome and decrease the sus- ceptability to cell death. Furthermore, overexpression of Bc12 protects these abnormal cells from apoptosis. All these derangements of cell cycle control should contribute to the human cervical carcinogenesis.

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Relationship between Expressions of Tumor - Associated Antigen MAGE-3 and p53 Proteins during Cell Cycle by Bivariate Analysis of Flow Cytometry

Hee Kyoung Chang, Deok Jun Kim, Kang Dae Lee, Hwan Jung Roh, G Spagnoli

J Korean Cancer Assoc. 1999;31(4):784-792.

Abstract PDF

PURPOSE
MAGE (melanoma antigen gene) is a tumor associated antigen, presented by HLA class I molecules, which is recognized by cytotoxic T lymphocytes. The expression of MAGE proteins are confined to malignant tumor tissues, except for the normal testis and placental tissues. Therefore, MAGE may be a potential target for immunotherapy of malignant tumors. However, biological aspects associated with the cell cycle are not yet described.
MATERIALS AND METHODS
The material used for this study was a novel human squamous cell carcinoma cell line (PNUH-12) from the hypopharynx, which had one point mutation of 78th base, C to G, in exon 7 of p53 gene. To understand the role of MAGE in relation to cell cycle and its relationship with p53 as the Gl checkpoint regulator, the expressions of MAGE-3 protein and mvtant p53 (Mtp53) were accessed by flow cytometry and immunohistochemistry. Double stains for MAGE-3/Mtp53 was analyzed with bivariate flow cytometry. DNA histograms using MAGE-3/PI (DNA) and Mtp53/PI (DNA) were also analyzed.
RESULTS
The expression rate of MAGE-3 and Mtp53 were 83% and 85%, respectively. MAGE-3 was expressed in cytoplasm, while M:p53 were expressed in the nuclei of the tumor cells on the immunohistochemical sections. With bivariate analyses, coexpression rate of MAGE-3/Mtp53 was 0.96, and MAGE-3 and Mtp53 constantly showed high levels throughout the cell cycle except Go.
CONCLUSIONS
These results mean that (I) MAGE-3 might have yet unknown relationship with mutant p53, (2) expressions of MAGE-3 and Mtp53 are not dependent on the cell cycle in PNUH-12 hypopharyngeal squamous carcinoma cell line, and suggest that MAGE-3 might have a role as important as p53 during the development of malignant tumors.

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Chromosomal Alterations in Gastric Cancer Cell Lines Detected by Comparative Genomic Hybridization

Mahn Joon Ha, Sang Jin Park, Hyun Woong Kang, Hyon Ju Kim, Hugh Chul Kim

J Korean Cancer Assoc. 1999;31(2):219-229.

Abstract PDF

PURPOSE
There are only a few cytogenetic studies in gastric cancer and so far no consistent specific chromosomal abnormalities have been described. In this study, we have used comparative genomic hybridization (CGH), a powerful molecular cytogenetic technique for detecting changes of the copy number throughout the genome, to screen for genetic alterations in gastric cancer cell lines. The CGH results were compared with those derived from G-banding and chromosome painting.
MATERIALS AND METHODS
Conventional cytogenetic analysis was performed on five human gastric cancer cell lines, AGS, SNU-1, SNU-16, SNU-620, and SNU-719, by a G-banding staining technique. In CGH, equal amounts of differently labeled DNA from the cell lines and normal reference DNA were hybridized simultaneously to normal metaphase chromosomes. They were visualized by different fluorochromes, and the signal intensities were quantitated separately as gray levels along the single chromosomes. The over- and under- represented DNA segments were determined by computation of ratio images and average ratio profiles. To confirm the CGH results, fluorescence in situ hybridization (FISH) with chromosome specific painting was performed using indirectly labeled chromosome specific paints.
RESULTS
Complex unbalanced chromosomal aberrations that could not be identified reliably by conventional cytogenetics in gastric cancer cell lines were successfully resolved by CGH analysis. CGH results were validated by using FISH with chromosome specific probes. In gastric cancer cell lines, gains of DNA copy number were more common than losses. Gains were detected on 1p, 1q, 2p, 3q, 6p, 7q, 10q, 11p, and 19q, and losses were observed on 4p, 4q, 5q, 12p, 12q, and 18q. Interestingly, all the five gastric cancer cell lines tested showed gain of DNA copy number on the chromosome 20, suggesting an existence of oncogene.
CONCLUSION
Conventional cytogenetics, CGH, and FISH using painting probes represent complementary approaches that, when employed in combination, could greatly facilitate the comprehensive analysis of chromosomal imbalances in gastric cancer cell lines. Our results suggest the existence of an oncogene or oncogenes on chromosome 20 that play a role in the development and/or the progression of gastric carcinogenesis.

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The p16INK4A Expression in Stomach Cancer , Colon Cancer and Hepatoma Cell Lines

Sun Ju Choi, Soo Kie Kim, Se Jong Kim, Choon Myung Koh, Yoon Sun Park

J Korean Cancer Assoc. 1998;30(3):527-535.

Abstract PDF

PURPOSE
The p16(INK4A) gene encodes a specific inhibitor of cell cycle progression. In recent years, genetic deletion and altered expression of p16(INK4A) gene were frequently showed in many human cancers. So, the p16(INK4A) gene is considered as tumor suppressor gene. However, there has been a few data for the p16(INK4A) in gastric cancer, colon cancer, and hepatoma.So.we investigated the genetic deldtion and altered expression of p16(INK4A) in gastric cancer, colon cancer and hepatoma cell lines.
MATERIALS AND METHODS
The homozygous deletion of p16(INK4A) was examined by using PCR and the protein expression of p16(INK4A) by using Western blotting in cancer cell lines established from Korean patients: stomach cancer, colon cancer and hepatoma cell lines.
RESULTS
Homozygous deletion of p16(INK4A) was detected only 1 stomach cancer cell line out of 13 cell lines examined. The p16(INK4A) was detected in 3 of 13 cancer cell line. These results showed the low frequency of p16(INK4A) homozygous deletion and high frequency of p16(INK4A) expression alteration in stomach cancer, colon cancer and hepatoma cell lines.
CONCLUSION
In this study, it may be suggested that the altered pl6(INK4A) expression as well as p16(INK4A) gene deletion play important role in oncogenesis. Further studies to determine the mechanism of p16(INK4A) gene inactivation are expected.

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A Study on the Tropism of Epstein-Barr Virus

Choon Hae Chung, Mi Ja Lee, Ho Jong Jeon

J Korean Cancer Assoc. 1997;29(6):954-964.

Abstract PDF

PURPOSE
The Epstein-Barr Virus (EBV) is associated with a variety of human lymphocytic and epithelial malignancies. EBV is thought to display exclusive tropism for B lymphocytes, follicular dendritic cells, and pharyngeal epithelia via specific receptors (C3d receptors, CR2, CD21). Recent evidence, however, challenged this belief. We designed this experiment to determine the incidence of EBV receptor in various malignant tumor cell lines and normal lymphocyte subsets.
MATERIALS AND METHODS
We have examined the incidence of EBV receptor, CD21 on the 10 healthy adult peripheral blood (PB), 10 umbilical cord blood (CB), 4 immortalized lymphoblastoid B cells by EBV infection (CSUP-1, CSUP-2, CSUP-3, CSUP-4), 3 EBV-positive B cell lymphoma cell lines (Jiyoye, IM-9, PTLC-1), 1 EBV-negative B cell lymphoma cell line (JeKo-1), 3 T cell lymphoma and leukemia cell lines (CCRF-CEM, H9, CEM-CM3), one histiocytic lymphoma cell line (U-937) and 5 gastric cancer cell lines (KATO III, AGS, SNU-1, SNU-5, and SNU-16). EBV receptor, C3d receptor was identified by flow cytometry (FACSCalibur) using FITC-conjugated or PE-conjugated CD21 monoclonal antibody. Also we investigated the expression of CD3, CD5, CD7, CD19, CD20, IgM, IgG, Ig and Ig by using FITC-conjugated or PE-conjugated monoclonal antibody, on above cell lines.
RESULTS
The expressions of CD21 molecule were 10.99 3.84% and 9.22 5.39% in adult PB lymphocytes and CB lymphocytes, respectively. The anti-human CD21 antibody was positive for CD19-positive or CD20-positive B lymphocytes. The CD3-positive or CD7-positive T lymphocytes were negative for anti-human CD21 antibody in PB and CB. But, CD21 antibody was weakly positive for CD5-positive lymphocytes. EBV-positive cell lines expressed variable ranges from 0.9% to 5.2% for CD21 antigen, while EBV-negative lymphoma cell line, JeKo-1 expressed 5.5%. All T lymphoma and leukemia cell lines and gastric cancer cell lines did not express CD21 antigen. But U-937 expressed 14.4% for CD21 antigen.
CONCLUSION
These results suggested that the CD21 antigen was expressed in CD20 or CD19-positive mature B cells, CD5-dim positive lymphocytes, some EBV-positive and negative B cell lymphoma cell lines, and a histiocytic lymphoma cell line. Further evaluation on the nature of CD5-dim positive cells, which was expressing CD21 molecule, is revealed, especially in reference to EBV association in some peculiar subtypes of peripheral T cell lymphoma.

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Alteration of Oleate-Phospholipase D Activities in Some Cell Lines after Irradiation

Chul Yong Kim, Myung Un Choi, Myung Sun Choi

J Korean Cancer Assoc. 1997;29(6):944-953.

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PURPOSE
Phospholipase D (PLD) catalyzes the hydrolytic cleavage of terminal phosphate diester bond of glycerophopholipids to produce phosphatidic acid (PA). PLD plays an important role in signal transduction and is known to be involved closely in cancer promotion, inflammation, and other cell responses. In order to evaluate radiation effect in tumor cells, various cells were screened for PLD activities and examined their radiation effects on PLD following gamma- ray irradiation.
MATERIALS AND METHODS
PLD activities in 19 species of cell were measured by radioactive isotope method with 1,2 - di [1-14C] phosphatidylcholine in the presence of oleate. Among the cell lines examined, VERO 76, L 1210 and P 388 were selected and examined for their effects of metal ions and agonists on PLD activities before and after irradiation by Co-60 teletheraphy unit.
RESULTS
The activities of oleate-PLD were observed in 11 species among 19 cell lines examined. VERO 76 and L 1210 cells showed that the PLD activity increased immediately after irradiation and reached to 150~200% of the control levels. The activation of PLD in response to gamma-ray was maximum at 20 Gy. In irradiated VERO 76, the stimulatory effect of Mg2+ was reduced and the activation of PLD by agonists in irradiated cells vary from those of the control cells.
CONCLUSION
The activation effect of irradiation on PLD activity observed strongly implies that the PLD activity is closely related to the phenomena of cell necrosis. Therefore the cell lines examined here could provide a good source for the study of radiobiology that cover from cell death to cancer promotion.

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Limited Cytotoxic Effect of Adenoviral-mediated p53 Gene Transfer in Variable Non-small Cell Lung Cancer (NSCLC) Cell Lines

Jhingook Kim, Sook Hyun Lee, Eun Sung Hwang, Jong Sik Kim, Kwhanmien Kim, Je Ho Lee

J Korean Cancer Assoc. 1997;29(4):565-575.

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PURPOSE
Cancer gene therapeutic strategy using p53 tumor suppresser gene have been suggested to be effective in many solid tumors including non-small cell lung cancer (NSCLC). To test generalized applicability, we tested a number of non-small cell lung cancer cell lines for their sensitivity to adenoviral-mediated wild-type p53 gene transfer.
MATERIALS AND METHOD
Replication-incompetent recombinant adenovirus encoding wild- type p53 (Avp53) under the control of the human cytomegalovirus (CMV) promoter was constructed and the cytotoxic effectiveness was evaluated in various NSCLC cell lines. Because 20 moi (multiplicity of infection; number of active virus particle/cell number) of Avp53 showed highly-effective cytotoxicity in p53-deleted cell lines (NCI-H1299, and NCI-H358), same amount was used for other cell lines.
RESULTS
Variable degree of cytotoxicity were observed in cell line with p53 mutation, but almost no effect were observed in those with will-type p53. Neither the infectivity of adenovirus, which was observed by x-gal stain after adenoviral mediated lac Z gene, nor the expression of p53 protein in infected cell, which was observed by western blot, was not the useful marker to expect the cytotoxic effect of Avp53. However, in responsive cell lines with Avp53, prominent expression of p21 protein, which was observed by western blot, was noticed.
CONCLUSION
In conclusion, adenoviral-mediated wild-type p53 transfer may not be applicable to every patient with non-small cell lung cancer, especially when the tumor has wild-type p53 gene. Better method to predict the effectiveness before application and strategy to widen the applicable extent is needed.

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Effects of Interleukin-2 Transduction into the Human Hepatoma Cell Lines Using Retroviral Vector

Soo Jung Gong, Nae Chun Yoo, Joo Hang Kim, Dong Hwan Shin, Hyo Dong Uhm, Sook Jung Jeong, Jae Yong Cho, Sun Young Rha, Yeon Soo Kim, Hyun Cheol Chung, Jae Kyung Roh, Jin Sik Min, Byung Soo Kim

J Korean Cancer Assoc. 1997;29(4):555-564.

Abstract PDF

PURPOSE
We compared the differences between parent hepatoma cell lines and interleukin-2 (IL-2) transduced hepatoma cell lines using N2A/IL-2 and LNC/IL-2 retrovirus with regards to in vitro sensitivity to peripheral blood monocytes and in vivo tumorigenic activity.
MATERIALS AND METHODS
Retroviral vector and producer cell line were constructed and IL-2 gene was transduced into the human hepatoma cell lines (SK-Hep1, Hep-G2, Hep-3B). IL-2 secretion after IL-2 transduction was measured by ELISA. MTT assay for in vitro sensitivity to peripheral blood monocytes was performed and the tumorigenic activity was observed in BALB/c mice and nude mice.
RESULTS
IL-2 secretion was 186 pg/10 degrees C cells/24 hrs in SK-Hep1 cell line and was 147 pg/10 (6) cells/24 hrs in Hep-3B cell line with N2A/IL-2 retroviral vector and was 55,000 pg/10 (6) cells/24 hrs with LNC/IL-2 retroviral vector. In vitro sensitivity to peripheral blood monocytes was increased by 163.8~254% in IL-2 transduced hepatoma cell lines (Hep -3B/N2A/IL-2, Hep-G2/N2A/IL-2) compared to those of the parent cell lines. The tumorigenicity was observed in 1 of 3 BALB/c mice and all 3 nude mice. Simultaneous injection of 1 X 10 (7) cells of the parent cell line (Hep-3B) into the right flank and IL-2 transduced cell line (Hep-3B/LNC/IL-2) into the left flank of the three BALB/c mice and of 5 X 10 (5) cells for the three nude mice resulted in a complete regression of the IL-2 modified tumor cell line (Hep-3B/LNC/IL-2) in 3 weeks and the parent cell line (Hep-3B) in 5 weeks. But, after the injection of 1.5 X 10 (7) cells for other five nude mice, the tumor of the IL-2 transduced hepatoma cell line (Hep-3B/LNC/IL-2) was gradually disappeared, and the tumor of the parent hepatoma cell line (Hep-3B) was initially decreased and then gradually regrew 20 days later.
CONCLUSION
IL-2 transduced hepatoma cell lines secreting IL-2 became more sensitive to peripheral blood monocytes and resulted in the increased antigenicity to the tumors formed by IL-2 transduced hepatoma cell line and parent cell line, and finally resulted in the regression of the tumors in experimental animals.

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Expression and Intracellular Localization of Hepatitis C Viral Core Protein in Human Hepatoma Cell Line Transfected with Viral cDNA

Dong Wan Kim, Sun Hee Kim

J Korean Cancer Assoc. 1997;29(1):11-18.

Abstract PDF

PURPOSE
Establishment of the human liver cell lines which permanently express the HCV proteins is important for the large scale production of viral antigen and analysis of the mechanism of hepatocellular carcinogenesis by HCV. Here, we attempted to establish the human hepatoblastoma cell lines which stably express the HCV core protein and examined the intracellular localization of the core protein.
MATERIALS AND METHODS
The cDNA of HCV core protein and neomycin resistance gene were expressed in HepG2 cells by the SRalpha promoter and human EF-1alpha gene promoter, respectively. The core protein was detected by immunofluorescence assay and western blotting.
RESULTS
We obtained several HepG2 cell clones which express HCV core protein stably. In transient expression assay, the core protein was localized in the cytoplasm in about 90%, and localized in the nucleus in about 10% of the core-expressing HepG2 cells. But, in the stably expressing HepG2 cell clones, the core protein was localized only in the cytoplasm. No HepG2 cell containing core protein in the nucleus was found in all of the cells which stably express the core protein.
CONCLUSION
The EF-1alpha gene promoter is highly efficient in the colony formation by neomycin resistance gene and is very useful for the isolation of human liver cell clones which express foreign genes stably. HCV core protein is localized in both nuclear and cytoplasm of human liver cell in short term but the cells containing the core protein in nucleus seem to disappear in long term culture.

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An Experimental Study on In Vitro Chemosensitivity Tests Using Human Cancer Cell Lines

Dae Hyun Yang, Jin Pok Kim

J Korean Cancer Assoc. 1989;21(2):376-406.

Abstract PDF

The more effective chemosensitivity tests should be developed as useful tools in cancer biology research, anticancer drug screening, individualized chemotherapy, and development of other cancer treatment modalities. For practical use of chemosensitivity tests enormous efforts are necessary to improve previous tests or to develop new ideal tests. In this study, the experiments to achieve more efficient experimental conditions such as proper plating cell numbers and reasonable anticancer drug exposure (concentration and timel in the clonogenic assay using 5 human cancer cell lines were performed, and the inhibitory effects of anticancer agents on in vitro tumor marker levels of 2 cancer cell lines were evaluated, and then the value of simple dye exclusion assay was reassessed. The human cancer cell lines used in this study were SC-I of colon cancer, PLC/PRF/5 of hepatoma. ZR-75-1 of breast cancer, G 361 and Bowes melanoma of malignant melanoma. The anticancer drugs used were 5 FU, methotrexate, mitomycin C, adriamycin, cisplatin and BCUN. The clonogenic assay of SC-1 was done at various plating cell numbers, and clonogenic assay as chemosensitivity test was performed at various drug concentrations with continuous or 1-hour exposures. Clonogenic assays of 5 human cancer cell lines were carried out at the drug concentrations of 1/10 peak plasma concentra- tion (PPC) and 1/100 PYC of continuous exposure, or 1/1(l PPC of I-hour exposure. The tumnor marker inhibition of SC-I and PLC/RRF/5 by anticancer drug was measured by radioimmunoassay on the third and the fifth incubation day, and the cell survival fraction by dye exclusion assay with trypan blue was calculated. In clonogenic assay of SC-I; the number of colrmies was increased with the increase of plating cell number, and the piating of: 5 x 10(4) cells/mi formed the sufficient number of colonies (over 500 per well) with reasonable coefficiency of variance. Other cell lines also formed sufficient number of colonies with the plating of 5 x 10(4) cells/mi. In clonogenic assay of SCI as chemosensitivity test; continuous exposures of 1/10 PPC and 1/100 PPC of anticancer drug and 1-hour exposure of 1/10 PPC showed valuable results which were able to differentiate more sensitive drugs from less sensitive drugs. In clonogenic assay of 5 cancer cell lines: continuous exposures of 1/10 and 1/100 PPC identified sensitive drugs more clearly than 1-hour exposure of 1/10 PPC. Most anticancer drugs of long in vitro half life revealed more sensitive activity than those of short half life in the clonogenic assay of continuos exposure. Among three kinds of in vitro chemosensitivity tests of continuous drug exposure, the clonogenic assay showed more sensitive data in anticancer activity than the tumor marker inhibition test and the dye exclusion assay. And the results of 3 tests were closely interrelated in many series of the experiment. Anticancer drugs of longer in vitro half life usually revealed more sensitive activity also in the tumor marker inhibition test and the dye exclusion assay of continuous exposure. In conclusion: in the experiments of in vitro chemasensitivity tests using human cancer cell lines, the proper plating numbers of the clonogenic assay were approximately 5x10(4) cells/ml, and the continuous drug exposures (1/10 PPC and 1/100 PPC) identified sensitive drugs more clearly than the 1-hour exposure (1/10 PPC), and the effects of the exposure time and especially the in vitro drug half life in media as well as the in vitro anticancer drug concentration were significant on the sensitivity data in the clonogenic assay and also in the tumor marker inhibition test and the dye exclusion assay, and the results of three tests were closely interrelated in many series of the experiment.

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The Combined Effects of 5 - Fluorouracil and Recombinant Interferon - gamma on Human gastric Cancinoma Cell Lines

Heung Tae Kim, Jae Gahb Park, Jin Pok Kim, Seong Hoe Park, Noe Kyeong Kim

J Korean Cancer Assoc. 1990;22(3):458-476.

Abstract PDF

Stomach cancer is a leading malignant disease in many countries. Conventional combination chernotherapy approaches to advanced gastric cancer only produce paitial response and there has been no impact on patient survival from these approaches as well. Of several promising new approaches the combination of interferon (IFN) and chemotherapeutic agents are now being made to improve the effectiveness for the treatment of cancer. Preclinical studies suggested that IFN may biochemically modulate the cellular uptake or metabolism of 5-fluorouracil (5-FU) resulting in s synergistic antitumor effect. Based on these data, Wadler reparted a promising result with combina- tion of 5-FU and IFN-alpha in patients with advanced colorectal carcinoma. This study was conducted to investigate the combined effects af 5-FU and recombinant IFN- gamma at cellular level against four gastric carcinoma cell lines (SNU-1, SNU-5, SNU-16, and NCI-NB7). We used a semiautomated tetrazolium-based colorimetric (MTT) assay for cytotoxicity and an isobologram analysis to evaluate the effect of the combination. The experiment was perfor- med three times on each of the three cell lines. Only two experirnents for SNU-16 and NCI-N87 showed supraadditivity (p< 0.02). On isobologram plotted by the mean value of three experiments for each cell line, supraadditivity was suggested for only SNU-16 (p = 0.055). In conclusion, our result did not document in vitro synergy between 5-FU and IFN-gamma for gastric carcinoma cell lines but additivity within clinically achievable dose range. Because in vivo immunomodulatory effect of IFN-gamma on host is more important rather than antiproliferative effect, the combination of 5-FU and IFN gamma is expected to improve the treatment of advanced gastric cancer.

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Significance of Serum Total Lactate Dehydrogenase ( LDH ) Level and Isoenzyma Patterns in Non - Hodgkin's Lymphoma as a Prognostic Factor

Jee Sook Hahn, Hyun Cheol Chung, Joo Hang Kim, Sun Ju Lee, Eun Hee Koh, Jae Kyung Roh, Hyon Suk Kim, Yun Woong Ko, Byung Soo Kim

J Korean Cancer Assoc. 1990;22(3):476-490.

Abstract PDF

Serum total lactate dehydrogenase (LDH) and LDH isoenzyme activities were studied in 33 cases w i th non-H odgk in's lymphom a to compare the cl inical significance of serological staging with these two serological markers to anatomical staging. Serum total LDH activity correlated with tumor burden as determined by clinical stage at presentation. Initially, LDH-3 was the major fraction of increment for the reflection of increased serum total LDH activity. With the increment of tumor burden (total LDH >200 IU/L), LDH-3 with additional LDH-4 fraction increased, which resulted in LDH-1/LDH-3 flipped pattern. These changes were normalized if complete remission state was induced with treatment. A high pretreatment LDH level (total LDH> 200 IU/L) correlated significantly to a decreased survival rate IP.0.01l> Furthermore, the flip pattern of LDH-I/LDH-3 isoenzyme at diagnosis showed a decreased survival rate (p<0.05), in which 4-year survival rate of patients with non-flipped pattern was 73.8%, comparable to 25.4%; of patients with flippd pattern. The 4-year survival rate of the low risk group Itotal LDH 200 IU/L with unftipped pattern: serological stage A) was 73g while 2-year survival rate of the high risk group (total LDH>200 IU/L withflipped pattern: serological stage D) was 0%, which showed a significant difference (p<0.05). Stepwise Cox regression analysis to identify the important prognostic factors among the serum total LDH, LDH1/LDH-3 flip, serological stage, anatomical stage, B symptoms, cell type, hepatos- plemomegaly, mediastinal mass reveated that the serological stage was the only prognostic factor. In conclusion, based on results of the multivariate analysis, we propose a new prognostic classification of patients with serological staging system in non-Hodgkins lymphoma. Furthermore, the reproducibility and therapeutic stratigies will be warranted.

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Synergistic Effect of Cisplatin and 5 - Fluorouracil Against Gastric and Colon Cancer Cell Line

Yoon Sik Kang, Dong Young Noh, Jae Gahb Park, Kuk Jin Choe

J Korean Cancer Assoc. 1990;22(3):490-505.

Abstract PDF

The purpose of this study was to elucidate the synergistic effect of cisplatin and 5-fluorouracil against human gastric and colon cancer cell lines. Since it is virtually impossible to document supraadditive tumor cell kill in vivo, this in vitro experiment implicates the interaction of these agents at a cellular level. We used an in vitro tetrazolium-based colorimetric assay for cytotoxicity (MTT assay) and an isobologram analysis to test combination effects of the two drugs against gastric and colon cancer cell lines, SNU-1, SNU-5, SNU-16, SNU-CI, SNU-C2A, SNU-C4, SNU-C5 those were established at cancer research institue of Seoul National University and NC1-N87 from NCI, VSA. As a resuh, those drug combination applied to this experiment didnt show a synergistic effect except in SNU-1 cell line demonstrated partly synergistic effect. Then we conclude that there are lack of in vitro synergistic effect between cisplatin and 5- fluorouracil against gastric and colon cancer cell lines, and that methodology we adopted can be one index ta check the combinatian effect of anticancer drugs.

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MDR1 Gene Expression and Chemosensitivity to Anticancer Drugs in SNU Hepatocellular Carcinoma Cell Lines

In Gyu Hong, Sung Gyu Lee, Hyun Ju Lee, Gyung Hwa Him, Hun Sik Kim, Jae Gahb Park

J Korean Cancer Assoc. 1994;26(2):219-231.

Abstract PDF

Using recently established 8 SNU human hepatocellular carcinoma cell lines and Hep 3B cell line, we performed in vitro chemosensitivity test using semiautomated tetrazolium-based colorimetric(MTT) assay. We also measured the MDRl gene expression of these cell lines by measuring MDR1 RNA level using slot blot analysis. IC values(concentration of chemotherapeutic agent which produces 50% growth inhibition of tested cells) were in the range of 1.92~5000 ↑ ug/ml for 5-fluorouracil, 0.24~32.00 ug/ml for doxorubicin, 0.16-10.29 pg/ml for mitomycin-c, 2.00~16.56 ug/ml for cisplatin, 1.15-182.32 ug/ml for etoposide, and 0.002 ↓L -24 ↑ug/ml for vincristine. Assay area under the curve(A-AUC) at IC: values showed that only A-AUC of VP-16 was in a clinically achievable range for only one of the 9 cell lines. All other drugs were out of the range of clinically achievabie AUC. Three cell lines(SNU-354, SNU-368, SNU-449) were MDR-positive. When these 3 cell lines were compared to MDR-negative cell lines, IC value for doxorubicin was significantly higher (P<0.05), suggesting that high expression of MDRl gene is responsible for doxorubicin resistance in hepatocellular carcinoma cell lines.

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Retroviral Vector - mediated Tumor Necrosis Factor - α Gene Transfer into Human Gastric Carcinoma Cell Lines

Jung Ae Rhee, Jung Ae Lee, Dae Seog Heo, Sung Koo Han, Noe Kyeong Kim

J Korean Cancer Assoc. 1994;26(5):677-688.

Abstract PDF

The tumor necrosis factor a (TNF-a) is a potent cytokine with antitumor activities including a direct cytotoxic effect on human cancer cells and the enhancement of a cytotoxic immune response against the tumor. However, its effectiveness in the human clinical trials is limited due to severe systemic toxicities. An alternative approach, that tumor cells are genetically engineered to secrete TNF-a locally to stimulate the immune system without systemic toxicities, is suggested as a form of gene therapy (tumor-cell-targeted lymphokine gene therapy). In this trial, cDNA encoding human TNF-a (TNF-NeoR) was introduced into five human gastric carcinoma cell lines using a retroviral vector to examine whether TNF-a gene could be transfected and expressed in gastric carcinoma cell lines in vitro. Successful transfer of TNF-a gene into five gastric csrcinoma cell lines was confirmed by polymerase chain reaction techniques. Supernatants (1: 2 dilution) from cultures of transduced gastric carcinoma cell lines demonstrated cytotoxicity to TNF-sensitive WEHI 164 cell lines in the range of 20-49%. TNF- transduced gastric carcinoma cell lines secreted TNF-a at the concetration of 479-8869 pg/10(6) cells-24 hours, whereas the parental cell lines did not secrete TNF-a. There were no differences in the growth rates between parental and TNF-transduced cell lines in vitro. The four TNF-transduced SNU cell lines showed the resistance to endogenous and exogenous TNF, except SNU-668 cell line. In conclusion, TNF-a gene was successfully transfected and expressed in gastric carcinoma cell lines in vitro. These data will be helpful in the development of tumor-cell-targeted lymphokine gene therapy for the treatment of advanced gastric carcinoma.

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Cyto - Melecular Conparison between Primary Squamous Cell Line from Bladder

Joong Shin Kang, Soo Sang Sohn, Dage Kwang Kim, Sung Ik Chang

J Korean Cancer Assoc. 1995;27(1):111-121.

Abstract PDF

To evaluate which cyto-molecular genetic evolutionary patterns take place during the in vitro establishment of permanent squamous cell carcinoma cell line of urinary bladder, cytomolecular genetic follow up was performed on primary culture for two weeks and on cancer cell line after contineued culture for three years in vitro. Near-diploid cells present on primary culture and near-hypertriploid or hypotetraploid cells, in contrast, present on cancer cell line. Chromosomal gains or losses are random from primary cancer to cell line. There are two kinds af structural cytogenetic abnormalities through progress in culture time. One is maintaining abnormal clone from original cancer to derived cancer cell line. Others are cytogenetic alteration during progessing culture time; Increasing and decreasing abnormal clones are coexisted in both grouy. Activation of oncogenes are different from primary cancer to cancer cell line. In conclusion, there are genetic alterations through progressing from primary cancer to cancer cell line. Due to these alterations, cancer ce11 1ines don't substituted for primary cancer and can not be availabe for using materials to choose production on monoclonal antibody and theraputic test.

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A Monocolnal Antibody P1 Against Leukocyte Common Antigen

Kyeong Cheon Jung, Sang Yoon Kim, Cheung Seog Park, Seong Hoe Park, Sang Kook Lee

J Korean Cancer Assoc. 1995;27(1):121-130.

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In Vitro Chemosensitivaty Test of SK-302 on Human Gastric Carcinoma Cell Lines

Soo Kie Kim, Woon Seob Shin, Yoon Sun Park, Sun Ju Choi, Kyung Ho Lee, Joo Young Park, Choon Myung Koh, Chan Mug Ahn, Weon Sub Park

J Korean Cancer Assoc. 1995;27(5):703-711.

Abstract PDF

SK-30ZB with quinomycin-related structure is considered to be potent antitumor antibiotic. There were several reports that quinomycins and their derivatives showed wide spectrum of anti-tumor activity. But there were no reports on anti-gastric tumor activity. We fould SK- 302B with anti-gastric tumor activity in vitro using our established anti-tumorsubstance screening system. Recent attention has been focused on the possibility of predicting the effectiveness of anti-cancer drugs on individual tumor through chemosensitivity tests. However, optimal conditions for in vitro chemosensitivity test on human gastric carcinoma cell lines were not well established. We attempted to establish optimal conditions and to evaluate compara- tively in vitro tumor cell cytotoxicity between SK-302B and adriamycin, using 7 human gastric carcinoma celi lines. The optimal cell number and culture duration for in vitro tumor cell cytotoxicity(TCC) and colony formation inhibition assay(CFIA) is 5x10(3) - 1x10(4) cells/well (TCC) and 2.510(3) - 10(3) cells/well(CFIA), 4 days(TCC) and 10 - l4 days(CFIA), respectively, for all gestric cancer cell lines. Under these conditions, we performed chemosensitivity test. Measurement of cytotoxicity(IC50) and colony forming efficiency revealed higher tumoricidal activity of SK-302 B against all tested gastric cancer cell lines than compared to that of adriamycin. In conclusion, we preseneted optimal conditions for in vitro chemosensitivity test on human gastric carcinoma cell lines. Using optimal conditions of this sytem, it could be demonstrated that SK-302B exerted a potent gastric cancer cell growth inhibition in vitro. Therefor, These data suggest that SK-302B may have a possibility of development as promising candidate of anti-gastric cancer chemotherapeutic agent.

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The Cytotoxic Activity of Sterol Derivatives from Pulsatilla Chinensis Regal

Jin Won Hyun, Yong Man Yang, Min Sook Sung, Ha Sook Chang, Woo Hun Paik, Sam Sik Kang, Jae Gahb Park

J Korean Cancer Assoc. 1996;28(1):145-151.

Abstract PDF

In order to search for antineoplastic components from plants, cytotoxic activity against human carcinoma cell lines was measured in 266 extracts from 236 plants using the MTT (3-[4,5-dimethyl thiazol-2-yl]-2,5-diphenyl tetrazolium bromide) method. As a result, the root of Pulsatilla chinensis (Ranunculaceae), which has been used as an insecticide in oriental medicine, showed a cytotoxic effect. Its structure was determined to be a mixture of ￥a-sitosterol-3-O-glucoside and stigmasterol-3-O-glucoside (3:1). Its IC value was 26￥ig/ml against SNU-I stomach cancer cell line.

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Studies on the pathological Changes of Human Hepatocellular Carcinomase and Their Cell Lines Induced by Adriamycin

Sang Yong Song, Chul Woo Kim, Woo Ho Kim, Chong Jai Kim, Gyeong Hoon Kang, Yong Il Kim

J Korean Cancer Assoc. 1996;28(4):611-622.

Abstract PDF

Through our study on morphologic changes occurring in hepatocellular carcinomas (HCCs) treated with trans-arterial chemo-embolization(TAE), we noticed that significant differences of morphologic features and proliferative activity existed between HCCs before and after TAE. In order to investigate whether such changes could be explained by adriamycin, we performed in vitro administration of adriamycin to HCC cell lines. Using three HCC cell lines(SNU-354, SNU-398, SNU-449) established in our hospital, we added 20 ￥ig of adriamycin to the culture dishes at the beginning and doubled the dose every three days. Three cell lines showed different drug susceptibility and SNU-398 was the most sensitive one. Initial morphologic changes were cellular edema and clearing. Damaged cells showed decrease of the size of cytoplasm and the cell processes became thinner and longer. Ultrastructurally, treated cells had small cytoplasm and scanty organelles. Some of the cells contained many lipid vacuoles. Nuclear chromatins demonstrated clumping tendency in the treated cell lines. Adriamycin caused shortening of doubling time in SNU-449, which was statistically not significant. No difference of ploidy was seen between untreated and treated cell lines. In summary, the morphological changes of hepatocellular carcinoma cell line induced by adriamycin in vitro is not exactly the same pattern as that of in vivo chemo-embolization procedure. The above findings suggested that the morphological changes of hepatocellular carcinoma induced by adriamycin chemo-embolization was induced not only by chemotherapeutic drug but also by accompanying environmental factors such as hypoxic insult.

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