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Volume 32(3); June 2000
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Original Articles
Expression of p53, p21/WAF1, bcl-2 and Loss of Heterozygosity for the Study of Apoptosis in Gastric Carcinoma
Hang Jong Yu, Joo Ho Lee, Woo Ho Kim, Kuk Jin Choe, Jin Pok Kim
J Korean Cancer Assoc. 2000;32(3):447-457.
AbstractAbstract PDF
PURPOSE
The purpose of this study was to correlate the immunohistdegrees Chemical expressions of p53, p21 and bcl-2, with their loss of heterozygosity (LOH) and clinical significance.
MATERIALS AND METHODS
Paraffin-embedded tissue sections from 30 patients with gastric car cinomas were examined for immunohistdegrees Chemical staining and LOH study. Primary antibodies used in immunohistdegrees Chemical staining were mouse mondegrees Clonal antibody to human p53, p21/ WAF1 and bcl-2. For PCR-LOH assays, D6S271, D6S105, D18S386, TP53, D17S796, and D17S786 microsatellite markers were used.
RESULTS
The expression rates of p53, p21 and bcl-2 were 76.7%, 80% and 3.3%, respectively. The expression of p21 was correlated with lymph node metastasis. LOH were found in 20.8% at D6S271, 42.3% at D6S105, 31.6% at D18S386, 39.1% at TP53, 40.9% at D17S796, and 50.0% at D17S786. No correlation was found between the immunohistdegrees Chemical expression and the LOH in these gene sites.
CONCLUSION
p53 and p21 were detected in high rate, whereas bcl-2 expression rate was very low in gastric adendegrees Carcinoma. Of them, overexpression of p21 was correlated with the tumor progression. High incidence rate of LOH may play an important role in gastric carcinogenesis. These findings suggest that the effects on apoptosis and cell cycle by p53 and p21 were important in development and progression of gastric cancer.
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Expression of p21/WAF1/CIP1, p27/KIP1 and p57/KIP2 Proteins in Gastric Adendegrees Carcinoma
Young Euy Park, Young Hee Choi, Kyoung Chan Choi, Seoung Wan Chae, Hyung Sik Shin
J Korean Cancer Assoc. 2000;32(3):457-466.
AbstractAbstract PDF
PURPOSE
p21/WAF1/CIP1, p27/KIP1 and p57/KIP2 are negative regulators of the cell division cycle. We evaluated the expressions of KIP CDK inhibitors and examined the relationship of clinicopathologic parameters and cell proliferation index in gastric adendegrees Carcinomas.
MATERIALS AND METHODS
The study was carried by the TUNEL method for apoptosis, immuno histdegrees Chemical staining for PCNA, p53, p21/WAF1/CIP1, p27/KIP1, and p57/KIP2 proteins and Western blot for KIPs proteins of normal and cancer tissues of stomach.
RESULTS
In normal gastric mucosa, p21/WAF1/CIP1 and p27/KIP1 proteins were expressed both mainly to the superficial portion of the glands with intestinal metaplasia and stromal cells. p57/KIP2 protein was also expressed normal crypt glandular epithelial and stromal cells. In gastric adendegrees Carcinoma, p21/WAF1/CIP1 was positive in 25 of 70 (35%) and showed significant decrease in deep tumor invasion (p=0.015) and the presence of angioinvasion (p=0.013). There was signi ficant inverse correlation between p27/KIP1 expression and cell proliferating index. p21/WAF1/ CIP1 expression was related to lower apoptotic index. Western blot analysis of KIP CDK inhibitors showed marked down-regulation of p21/WAF1/CIP1 protein in cancer than normal tissue, but alternative expression in p27/KIP1 and p57/KIP2 proteins.
CONCLUSION
The above results indicated that the expression loss of KIP CDK inhibitors was contributed to tumor progression and aggressiveness in gastric adendegrees Carcinoma.
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Amplification and Overexpression of c-erbB-2 in Gastric Cancer
Si Youl Jun, Ho Young Chung, Seung Cheol Lee, Tae Lin Huh, Wansik Yu
J Korean Cancer Assoc. 2000;32(3):467-475.
AbstractAbstract PDF
PURPOSE
We compared c-erbB-2 oncogene amplification and oncoprotein expression, trying to identify the biologic and prognostic significance of c-erbB-2 in adendegrees Carcinoma of the stomach.
MATERIALS AND METHODS
Formalin-fixed, paraffin-embedded tissue sections from 43 cases of gastric cancer were analyzed for amplification of c-erbB-2 by differential polymerase chain reaction and for overexpression of gene product by immunohistdegrees Chemistry.
RESULTS
The amplification was detected in 13 cases (30%). Enhanced c-erbB-2 immunoreactivity was observed in 30% (13/43) of tumors. Tumors with gene amplification generally stained strongly (p=0.003). Although the frequency of amplification and overexpression of c-erbB-2 was increased with advanced gastric cancer and with lymph node metastasis, this difference was not statistically significant. c-erbB-2 gene amplification or protein overexpression showed a trend toward a better five year survival rate, but this did not reach a statistical significance.
CONCLUSION
Amplification and/or overexpression of the c-erbB-2 may be of value in clarifying the biologic characteristics of the human gastric cancer. However, more sensitive and more speci fic methods of identifying gene amplification are needed and the standardization of the staining method as well as guidelines for interpreting the staining result are mandatory for this purpose.
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Expression of MDM-2 and p53 Proteins in Gastric Adendegrees Carcinoma and Its Relationship with Clinicopathologic Factors
Ji Woong Yang, Hyoung Joong Kim, Tae Jin Lee, Eon Sub Park, Jae Hyoung Yoo
J Korean Cancer Assoc. 2000;32(3):476-486.
AbstractAbstract PDF
PURPOSE
MDM-2 is an oncoprotein that inhibits p53 tumor-suppressor protein. These abnor malities have a role in tumorigenesis through inactivation of p53 function. To determine the clini copathological and prognostic value of MDM2 abnormalities in gastric adendegrees Carcinoma, MDM-2& p53 protein expression were analysed in surgically resected materials of gastric adendegrees Carcinoma.
MATERIALS AND METHODS
Fifty cases which had got follow-up after surgical resection were immunohistdegrees Chemically studied with p53 and MDM-2 antibodies. We defined variable clinico pathologic factors for expression of p53 and MDM-2 protein and analysed their relationships.
RESULTS
Immunohistdegrees Chemical stain revealed expression of MDM-2 protein as a 52.0% (26/50) and p53 protein 20.0% (10/50), respectively. But their expressions were not assdegrees Ciated with clinicopathological factors such as T-factor, N-factor, stage, histology and differentiation. Overall, p53-negative patients seemed to have a better prognosis regardless of MDM-2 protein status (P= 0.057). MDM-2 protein status was considered to have no play as a prognostic factor.
CONCLUSION
In the gastric adendegrees Carcinoma, p53 protein expression seemed to have a inverse relationship with clinical outcomes but MDM-2 protein expression, which was observed more frequently than those of p53, seemed not to be prognostic indicator.
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Clinicopathologic Significance of Multidrug Resistance Protein Expression in Patients with Stage III Gastric Adendegrees Carcinoma
Sae Hyun Kim, Wan Ku Lee, Hyun Kim, Young Nam Kim, Seung Min Park, Su Jin Choi, Hyo Suk Park, Myung Jin Joo, Kwang Min Lee, Jong Myung Lee, Sung Hye Shin, Min Chul Kim
J Korean Cancer Assoc. 2000;32(3):487-496.
AbstractAbstract PDF
PURPOSE
We wanted to determine the prognostic significance of P-glycoprotein (Pgp) and multi drug resistance-assdegrees Ciated protein (MRP) in stage III gastric adendegrees Carcinoma by evaluating whe ther the Pgp and/or MRP expression correlate with various clinicopathological parameters and survival rates. MATERIAL AND METHODS: The expression of Pgp and/or MRP were studied immunohistdegrees Chemi cally by ABC method with paraffin-embedded tissue specimens which were surgically obtained from 64 cases of stage III gastric adendegrees Carcinomas at the Department of Surgery, Presbyterian Medical Center from 1991 to 1992. Statistical differences of both expression in various factors including survival rates and clinicopatholgical parameters were sought.
RESULTS
Expression rates of Pgp and MRP group were 50.0% and 43.7% respectively. There was no significant correlation between expression of two proteins and various clinicopathological variables such as age, sex, stage, tumor depth, number of metastatic node, tumor size, site and method of operation. However, in case of the degree of differenciation, the expression of Pgp and/or MRP was significantly greater in well differenciated adendegrees Carcinoma than in poorly dif ferenciated adendegrees Carcinoma (p=0.001, p=0.012). Statistically, no significant correlations between the expression of Pgp and/or MRP and overall survival rates were found.
CONCLUSION
These results suggest that the Pgp and/or MRP expression in patients with stage III gastric adendegrees Carcinomas are not useful in determining postoperative chemotherapy and as an independent predictor of survival.
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Interrelation between the Progression of Gastric Adendegrees Carcinoma and the Matrix Metalloproteinases
Sung Chul Lim, Mi Ja Lee, Chan Kook Park
J Korean Cancer Assoc. 2000;32(3):497-505.
AbstractAbstract PDF
PURPOSE
An essential element in tumor invasion and metastasis in the degradation of extracellular matrix is matrix metalloproteinases (MMPs). But little is known about the role of them in the gastric adendegrees Carcinoma. The purpose of this study was to determine whether human gastric adendegrees Car cinoma expressed MMPs and whether MMPs were concerned in tumor stage and histologic subtypes.
MATERIALS AND METHODS
The authors performed the retroactive analysis in 38 cases of early gastric carcinoma (EGC) and 61 cases of advanced gastric carcinoma (AGC), including 38 cases of diffuse type and 61 cases of intestinal type, using immunohistdegrees Chemical staining for MMP-2, -3 and -9.
RESULTS
MMP-2, -3 and -9 were ldegrees Calized to neoplastic cells in 93% (92/99), 32% (32/99) and 76% (76/99), respectively. MMP-2 and -9 were found mainly in tumors of intestinal type and less frequently in those of diffuse type. There were positive correlation between the presence of MMP-2 and -9 and lymph node metastasis. There was positive correlation between MMP-2 expression and tumor stage.
CONCLUSION
The authors demonstrated a statistically significant expression of tumoral MMP-2, -3 and -9 in gastric adendegrees Carcinoma and their correlation with tumor progression. This study also suggests that the presence of MMP-2 and/or -9 is a useful indicator for poor prognosis.
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Retrospective Analysis of Treatment Results of Recurrent Gastric Cancer
Hyuk Joon Lee, Sam Je Cho, Han Kwang Yang, Kuhn Uk Lee, Kuk Jin Choe, Jin Pok Kim
J Korean Cancer Assoc. 2000;32(3):506-515.
AbstractAbstract PDF
PURPOSE
Recurrent gastric cancer has a very poor prognosis due to its diagnostic difficulties, variant recurrence patterns and no effective treatment modalities. In this study, we retrospectively analyzed treatment results of the recurrent gastric cancer.
MATERIALS AND METHODS
We reviewed 1,286 patients who had taken radical surgery for primary gastric cancers and were diagnosed to have recurrences of their primary diseases. According to the medical records, we retrospectively analyzed the gross features, histologic types and TNM stages of primary gastric cancers. The symptoms, diagnostic modalities, durations, recurrence patterns, treatments and prognoses of recurrent diseases were also reviewed.
RESULTS
The median survival time of total recurrent gastric cancer patients was 6.8 months. Ldegrees Co-regional and distant recurrences had better results than peritoneal and mixed recurrences (p<0.01). Curative resection of recurrent cancer was done in only 1.6% (18 cases in ldegrees Co-regional recurrence and 4 cases in distant recurrence), but in these cases, significant survival gain (5 year survival rate: 30.5%) was found compared to other treatment modalities (p<0.01). In multivariate analysis, the type of first operation, TNM stage, duration till recurrence, recurrence pattern, and treatment modality of recurrence were meaningful for the survival time after recurrence.
CONCLUSION
In selective cases, recurrent gastric cancer had an effective treatment modality and was expected to have prolonged survival. Therefore, a careful diagnosis and an active treatment of the patients who have the recurrent gastric cancers should be done, especially for the ldegrees Co-regional recurrence.
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Chemotherapy with Five-Day Continuous Infusion of 5-Fluorouracil (5-FU) Plus Cisplatin for Advanced Gastric Cancer; Significance of 5-FU Concentration Monitoring
Yeon Hee Park, Bong Seog Kim, Baek Yeol Ryoo, Tae You Kim, Young Hyuck Im, Ho Sang Shin, Yoon Koo Kang
J Korean Cancer Assoc. 2000;32(3):516-523.
AbstractAbstract PDF
PURPOSE
To investigate the therapeutic effects and toxicities of 5-day continuous infusion of 5-FU plus cisplatin FP chemotherapy in advanced gastric adendegrees Carcinoma and to elucidate the relationship between the pharmacokinetic (PK) parameters and therapeutic outcome.
MATERIALS AND METHODS
Patients with previously untreated advanced stomach cancer were treated with FP chemotherapy. Plasma concentrations of 5-FU were measured using gas chro matography method for 5 days. Correlation of PK parameters of 5-FU with clinical outcome after FP chemotherapy was studied.
RESULTS
Response rate of FP chemotherapy was 46% (95% C.I.: 30~62%). There was a wide range of difference in the concentration and area under the curve (AUC) of 5-FU from patient to patient. We could find significant differences in AUC of 5-FU between the responders and the non-responders (p<0.05).
CONCLUSION
We could confirm that FP chemotherapy was effective and tolerable for the treatment of advanced stomach cancer. The monitoring of plasma 5-FU concentration after chemotherapy and the adjustment of subsequent 5-FU dose seems to be necessary to improve the treatment outcome of FP chemotherapy.
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Combination Chemotherapy with Etoposide, Doxorubicin, Cisplatin (EAP) for Recurred or Metastatic Gastric
Nam Kuk Cho, Tae Sik Choi, Yeon Hee Park, Seung Chul Lee, Young Jin Yuh, Sung Rok Kim
J Korean Cancer Assoc. 2000;32(3):524-530.
AbstractAbstract PDF
PURPOSE
We performed this study to evaluate the efficacy and the safety of EAP regimen as a second line therapy for the recurred or metastatic gastric cancer unresponsive to 5-fluorouracil based chemotherapy.
MATERIALS AND METHODS
Recurred or metastatic gastric cancer patients unresponsive to 5- fluorouracil based regimen were entered into this trial. They were treated by EAP chemotherpy which consisted of etoposide 40 mg/m2, doxorubicin 15 mg/m2 and cisplatin 25 mg/m2 IV during 3 days each every 3 weeks.
RESULTS
From December 1994 to March 1998, Eighteen patients were enrolled in this protdegrees Col. Fourteen patients were evaluable for response. The overall response rate was 28.6% (95% CI: 11.7~56.7%). The median response duration was 21 weeks. The median survival for all enrolled patients was 28 weeks. The major toxicity was myelosuppression. Among total of 69 cycle che motherapy, WHO grade 3 or 4 granuldegrees Cytopenia and thrombdegrees Cytopenia were observed in 71.0% and 27.5%, respectively.
CONCLUSION
Second line therapy with EAP regimen was active for gastric cancer. Chemotherapy induced toxicities were moderate to severe.
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The Role of bcl-2 and p53 in Tamoxifen-Induced Apoptosis of Human Breast Cancer Cell Lines
Woo Chul Noh, Dong Young Noh, Yong Ho Ham, Chang Min Kim, Nam Sun Paik, Nan Mo Moon, Kuk Jin Choe
J Korean Cancer Assoc. 2000;32(3):531-538.
AbstractAbstract PDF
PURPOSE
Tamoxifen has been well known as an effective anti-tumor agent against breast cancer. The important role of bcl-2 and p53 proteins in tamoxifen-induced apoptosis of breast cancer cells has been suggested. However, the paradoxical fact that bcl-2 over-expression is assdegrees Ciated with better prognosis in clinic has not yet been clearly explained. To investigate this paradox, we analyzed the effect and dynamics of bcl-2 and p53 on the apoptosis after treatment of breast cancer cells with tamoxifen.
MATERIALS AND METHODS
The human breast cancer cell lines MCF-7 and MB MDA-468 were treated with 17-betaestradiol (E2) and tamoxifen.
RESULTS
Following tamoxifen treatment, MCF-7 cells underwent apoptosis accompanied by reduced bcl-2 expression. E2 pre-treatment led to the inhibition of tamoxifen-mediated apoptosis and bcl-2 down-regulation. When MB MDA-468 cells were treated with E2 or tamoxifen, bcl-2 and p53 protein expression did not change and apoptosis did not develop.
CONCLUSION
We observed that the down-regulation of bcl-2 by tamoxifen treatment can facilitate the apoptosis of breast cancer cells without p53 mutations. This finding was consistent with clinical experiences in which bcl-2 positive tumors were assdegrees Ciated with more indolent phenotypes in breast cancer.
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The Effect of Long-term Tamoxifen Therapy on Endometrium Evaluated by Transvaginal Sonography in Postmenopausal Women Undergone Breast Cancer Surgery
Jung Hoon Bae, Lee Su Kim, Jeong Jin Kim, Min Kyun Lim, Jong Hyun Kim, Young Min Woo, Man Chul Park, Dong Kun Kim, Chang Sig Choi, Sung Kim
J Korean Cancer Assoc. 2000;32(3):539-544.
AbstractAbstract PDF
PURPOSE
Tamoxifen is a non-steroidal antiestrogenic drug used mainly in the adjuvant therapy of breast cancer and it also has estrogenic effect to the endometrium. We investigated the effects of tamoxifen on endometrial thickening in postmenopausal women taking adjuvant tamoxifen therapy for breast cancer, and analyzed the correlation between sonographic findings, pathologic findings and duration of tamoxifen therapy.
MATERIALS AND METHODS
Forty three patients previously operated and being treated by tamoxifen since July 1995 to August 1999 were involved in this study. Control group was selected from patients of postmenopausal syndrome visiting postmenopausal clinic more than 2 years after menopause since January 1994 to August 1997. Endometrial thicknesses of breast cancer patients taking tamoxifen were measured twice a year for 2 years and then once a year by transvaginal ultrasonography. They were considered abnormal when those thicknesses were greater than 6 mm. Sonographic mearsurements of control patients were done once at the first visit and which were compared with those of breast cancer patients.
RESULTS
The mean endometrial thicknesses of breast cancer patients receiving tamoxifen therapy were 5.06 mm (4.86~5.21 mm) at 6th month, 5.1 mm (4.92~6.00 mm) at 12th month, 5.13 mm (4.89~5.32 mm) at 18th month, 5.15 mm (4.97~5.28 mm) at 24th month and 5.14 mm (4.96~5.21 mm) at 36th month. The mean thickness of control patients was 4.87 mm. The mean endometrial thickness of breast cancer patients and that of control patients were 5.07 mm and 4.87 mm respectively. Only one patient with stage I breast cancer showed endometrial thicknessof 6 mm during follow-up and endometrial biopsy showed atypical endometrial hyperplasia. Endometrial thickness significantly increased after 18 months of tamoxifen therapy, but the rate of increase was slow.
CONCLUSION
The endometrial thickness increased with duration of tamoxifen therapy, but the rate of increase was slow and seldom exceeded 6 mm. So we concluded the risk of endometrial cancer is low.
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Phase II Multicenter Trial of Paclitaxel for Metastatic Breast Cancer
Jae Ho Byun, Sung Soo Yoon, Chan Hyung Park, Sung Rok Kim, Si Young Kim, Hyo Jin Kim, Soo Mee Bang, Heung Moon Chang, Dae Seog Heo, Yung Jue Bang, Noe Kyeong Kim
J Korean Cancer Assoc. 2000;32(3):545-552.
AbstractAbstract PDF
PURPOSE
To evaluate the efficacy and safety of paclitaxel for metastatic breast cancer patients who had received previous chemotherapy, we have performed phase II multicenter trial between December 1997 and December 1998.
MATERIALS AND METHOD
Thirty patients were accrued to this study and paclitaxel was administered at 175 mg/m2 as a 3-hour intravenous infusion every 3 weeks until the progression of the disease.
RESULTS
Objective response were observed in 13 of 30 patients (43.3%). There were 1 complete response (3.3%) and 12 partial responses (40%). Especially, 50% (11/22) of patients who had received prior anthracycline-containing regimens for adjuvant or metastatic disease responded to paclitaxel. Responses were observed in all sites of metastatic disease. One hundred forty-nine cycles of treatment were administered, with a median of six cycles per patient. Grade III and IV toxicities included neutropenia (24%), elevated liver enzyme (10%), peripheral neuropathy (10%), arthralgia/myalgia (23%), and alopecia (87%). No significant hypersensitivity type reaction or cardiac arrhythmia were seen. Median duration of response was 7.2 months.
CONCLUSIONS
These results suggested that paclitaxel is active therapeutic agent in metastatic breast cancer patients and it can be safely administered by 3-hour intravenous infusion with premedication.
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Alterations of p15INK4B, p16INK4A and Methylthioadenosine Phosphorylase Gene in Korean Hepatdegrees Cellular Carcinoma
Ho Young Pyun, Jae We Cho, Won Ki Baik, Jong Wook Park, Jae Pok Park, Min Ho Suh, Seong Il Suh
J Korean Cancer Assoc. 2000;32(3):553-562.
AbstractAbstract PDF
PURPOSE
We analyzed the gene status of p16INK4A, p15INK4B and MTAP (methylthio adenosine phophorylase) in Korean hepatdegrees Cellular carcinoma (HCC) to investigate whether the inactivation of these genes participated in hepatdegrees Carcinogenesis, and evaluated MTAP-targeted chemotherapy in MTAP-deficient cell lines. MATERIAL AND METHODS: We examined eleven primary HCC and 8 SNU cell lines using PCR, Southern blot analysis, PCR-SSCP, DNA sequencing, methylation-specific PCR, Western blot analysis, MTT assay, and crystal violet staining.
RESULTS
Mutations or deletion of the p16INK4A, 15INK4B, and MTAP genes were rare, but methylation of the p16INK4A promoter region was common in HCC. The base alterations of 3' untranslated region of p16INK4A exon 3 were also detected in 3 samples. In SNU cells, p16INK4A was not detectable, when treated with demethylating agent, high levels of re-expressed p16INK4A protein were detected. In MTAP-targeted chemotherapy experiment, methylthioadeno sine (MTA) was able to rescue MTAP positive cell lines but not MTAP negative cell lines from growth inhibition by depletion of methionine and MTX treatment.
CONCLUSION
These results suggest that de novo methylation of the p16INK4A promoter region seems to play an important role in the pathogenesis of HCC. And treatment of MTX, combined with methionine depletion in the presence of MTA, may be a high selective treatment for MTAP negative HCC.
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Sequential Changes of Proliferative Fraction of Enzyme Altered Fdegrees Ci in Experimental Rat Hepatdegrees Carcinogenesis
Woo Ho Kim, Yun Sil Yoo, Yong Il Kim, Mi Sook Lee, Min Jae Lee, Ja June Jang
J Korean Cancer Assoc. 2000;32(3):563-570.
AbstractAbstract PDF
PURPOSE
The proliferative activity of cells in enzyme altered fdegrees Ci of the rat hepatoma model was measured by double immunohistdegrees Chemical staining methods using anti-bromodeoxyuridine (BrdU) and anti-glutathione S transferase of placental form (GST-P). The aim of this study was to compare the cell proliferative activity in GST-P positive altered fdegrees Ci and in negative fdegrees Ci.
MATERIALS AND METHODS
Eight-week-old male Sprague-Dawley rats were administered by 200 mg/kg diethylnitrosamine (DEN) intraperitoneally, and followed by 0.02% acetylaminofluorene (AAF)-containing diet for 4 weeks. One week after administration of AAF diet, two-thirds hepa tectomy was performed. Control animals were treated as same except for the omission of AAF in the diet. The rats were sacrified 0, 1, 3, 5, 7, 14 and 21 days after partial hepatectomy. The slices of liver were fixed in acetone, dehydrated in benzene and stained by peroxidase-anti peroxidase method against GST-P and by avidine-biotin peroxidase complex method against BrdU.
RESULTS
The area of the GST-P positive fdegrees Ci was increased during the experimental period. In the experimental group, the S-phase fraction in the fdegrees Ci remained high during the first week and was decreased thereafter. However, the GST-P negative area maintained a low S-phase cell frac tion throughout the experimental period.
CONCLUSION
These results suggest that hepatic cells in the enzyme altered fdegrees Ci may escape a suppressor effect of AAF in contrast to the normal cells in which their growth are inhibited by AAF.
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Radioresponse of Hepatdegrees Celluar Carcinoma Treatment of Lymph Node Metastasis
Jong Hoon Kim, Eun Kyung Choi, Young Wha Chung, Young Sang Lee, Dong Jin Seo
J Korean Cancer Assoc. 2000;32(3):571-577.
AbstractAbstract PDF
No abstract available.
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Determination of Chromosomal Alterations in Nasal NK/T-cell Lymphomas by DOP-PCR and Comparative Genomic Hybridization
Sang Jin Park, Mahn Joon Ha, Hyon Ju Kim, Kwang Hwa Park, Hyun Soo Kim, Woo Ik Yang, Hugh Chul Kim
J Korean Cancer Assoc. 2000;32(3):578-586.
AbstractAbstract PDF
PURPOSE
Because of difficulty of obtaining metaphase cells from tumor specimens, there are only a few cytogenetic studies in nasal NK/T-cell lymphomas, and so far no consistent specific chromosomal abnormalities have been described. In this study, we have used degenerate oligonucleotide primed PCR (DOP-PCR) and comparative genomic hybridization (CGH) to deter mine chromosomal alterations from 6 nasal NK/T-cell lymphoma tissues dissected from formalin- fixed paraffin-embedded slide sections.
MATERIALS AND METHODS
For the isolation of tumor DNA, four 7-micrometer-thick tissue sections from each sample were dewaxed and rehydrated, and areas of high tumor cell content (more than 60%) were dissected and pooled into a tube. Normal DNA was prepared from the peripheral blood of a healthy volunteer. Tumor DNA was labeled with biotin-16-dUTP by DOP-PCR and normal DNA was labeled with digoxigenin-dUTP using a nick translation kit. In CGH, equal amounts of differently labeled DNA from the tumors and normal reference DNA were hybridized simul taneously to normal metaphase chromosomes. They were visualized by different fluordegrees Chromes, and the signal intensities were quantitated separately as gray levels for each chromosome. The over- and underrepresented DNA segments were determined by computation of image ratios and average ratio profiles.
RESULTS
Our results show that gains of DNA copy number were more prevalence than DNA losses. The most commonly observed gains were mapped to chromosomal regions of 1p32.2 ter,19 and 20 in 4 of 6 cases (67%). The other frequent gains were found on chromosomes 12q in 3 of 6 cases. The most frequent loss was detected on 6q in 4 of 6 cases(67%), and less fre quently observed on 13q21.1 q34 and 13q14 q34.
CONCLUSION
These genomic changes found in specific chromosomal regions are likely to harbor genes of importance in nasal NK/T-cell lymphomagenesis, therefore such cytogenetic mapping of genomic imbalance may be of value for further molecular delineation of NK/T-cell lymphoma.
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Frequency of bcl-2/JH Rearrangement in Benign Lymphoid Hyperplasia
Young A Yoo, Seung Ho Lee, Mi Na Son, Zeung Kun Cho, Kun Choi, Jong Wook Choi, Sang Won Shin, Byung Soo Kim, Jun Suk Kim, In Sun Kim, Yeul Hong Kim
J Korean Cancer Assoc. 2000;32(3):587-594.
AbstractAbstract PDF
No abstract available.
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Production of Soluble VEGF Receptor Mutants for Inhibition of Angiogenesis
Soo Young Yun, Yong Kil Hong, Yoon Lee, Kwangsei Kim, Hoon Kyo Kim, Young Ae Joe
J Korean Cancer Assoc. 2000;32(3):595-604.
AbstractAbstract PDF
PURPOSE
Vascular endothelial growth factor (VEGF) is a potent angiogenic factor of many solid tumors, promoting vascularization and formation of metastases. In an attempt to generate effective VEGF inhibitors, the authors constructed the VEGF receptor mutants, expressed in E. coli and Sf9 insect cells, and examined their binding to VEGF.
MATERIALS AND METHODS
The cDNA fragment encoding FLT-1 extracellular domain was cloned from human umbilical vein endothelial (HUVE) cell total RNA using RT-PCR. PCR- subcloning was performed using this template, in order to generate the deletion mutants by introducing FLT-1 partial sequences into E.coli expression vector pET-21d and baculovirus transfer vactors, pBAC-1 and pBAC-3. Two mutant proteins from baculovirus-infected insect cells were purified by heparin sepharose chromatography and immobilized into nitrdegrees Cellulose membrane followed by 125I-VEGF binding assay.
RESULTS
Two mutant receptors, sFLT (1~7) and sFLT (2~4) expressed in E.coli appeared in inclusion body as insoluble proteins. The soluble mutant receptors were produced in low yield by baculovirus/insect cell expression system. Both immobilized mutant receptors, sFLT (1~7) and sFLT (2~4) were able to bind VEGF.
CONCLUSION
These results suggest that a small soluble mutant receptor, sFLT (2~4), as well as sFLT (1~7) may be used effectively for bldegrees Cking angiogenic function of VEGF.
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Construction of DNA Painting Probe for Transldegrees Cation Region of a Marker Chromosome by Chromosome Microdissection
Chan Sang Park, In Hwan Lee, In Jang Choi, Dae Kwang Kim
J Korean Cancer Assoc. 2000;32(3):605-611.
AbstractAbstract PDF
PURPOSE
Chromosome microdissection has been recommended as a technology to overcome the limited problems of conventional cytogenetic analysis and is a direct approach to isolate DNA from specific interesting region of chromosome. KUMA-1 cell line has a specific reserved chromosome abnormality during prdegrees Cess from primary cancer culture to continuous cell line development, der(2)t(2;?)(qter;?). So molecular analysis for transldegrees Cation region of der(2) may be helpful to understand pathogenesis of this primary cancer. The aim of this study was to develop painting probe for the transldegrees Cation region for molecular study in future about transldegrees Cation region of der(2) of KUMA-1 cell line.
MATERIALS AND METHODS
KUMA-1 cell line was derived from a squamous cell carcinoma of urinary bladder. The transldegrees Cation breakpoint region of der(2) appeared in KUMA-1 cell line was microdissected and dissected chromosome segments were amplified by PCR reaction. Fluorescent in situ hybridization was conducted on KUMA-1 metaphase cells with the probe generated from PCR product to confirm the construction of painting probe containing the transldegrees Cation breakpoint of der(2).
RESULTS
Painting probe was hybridized to the metaphase chromosome of KUMA-1 cell line and two fluorescent signals were mapped to the transldegrees Cation forming chromosomal region of der(2).
CONCLUSION
It was possible to construct the painting probe for the transldegrees Cation region of der (2) by chromosome microdissection.
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Efficacy of Combination Chemotherapy with Vinorelbine, Ifosfamide, and Cisplatin in Patients with Advanced Non-Small Cell Lung Cancer
Heung Moon Chang, Jung Ae Lee, Jin Seok Ahn, In Sook Woo, Young Iee Park, Jee Woong Son, Seung Joon Lee, Dong Kyu Kim, Eun Kyung Mo, Myung Jae Park, Myung Goo Lee, In Gyu Hyun, Ki Suck Jung, Young Suk Park
J Korean Cancer Assoc. 2000;32(3):612-618.
AbstractAbstract PDF
PURPOSE
To evaluate the efficacy and toxicity of combination chemotherapy with vinorelbine, ifosfamide, and cisplatin in patients with advanced non-small cell lung cancer.
MATERIALS AND METHODS
Patients with unresectable, pathologically proven non-small cell lung cancer who had no prior chemotherapy were eligible. Patients received vinorelbine (25 mg/m2, iv., D1 & 8), ifosfamide (1.5 g/m2, iv., D1-3 with mesna), and cisplatin (60 mg/m2, iv., D1). The treatment was repeated every 3 weeks.
RESULTS
Between degrees Ctober, 1997 and June, 1999, 26 patients were enrolled. Median age was 61. 1 patient had stage IIIA, 13 had stage IIIB, and 12 had stage IV. Patients with adendegrees Carcinoma were 15, squamous cell carcinoma were 11. Of 22 evaluable patients, objective responses were observed in 9 patients (response rate: 40.9%, CR: 1 (4.5%), PR 8 (36.4%)). Median duration of response was 48 weeks. Median overall survival was 52 weeks. Grade 3-4 leukopenia was observed in 10.2% of the 88 courses. There was 1 death related to febrile neutropenia. Non- hematologic toxicities were mild.
CONCLUSION
We concluded that combination chemotherapy with vinorelbine, ifosfamide, and cisplatin was effective and tolerable in patients with advanced non-small cell lung cancer, and phase III randomized trial is needed to compare this regimen to other cisplatin-based regimens.
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Regulation of Gene Expression in Murine Renal Cell Carcinoma Cells Growing in Ectopic or Orthotopic Organs of Syngenic Mice
Yoo Sun Jung, Kwang Sung Ahn, Jhingook Kim, Hyunah Lee, Sung Soo Yoon
J Korean Cancer Assoc. 2000;32(3):619-628.
AbstractAbstract PDF
PURPOSE
The biologic behavior of tumor cells is partially controlled by the microenvironment. We investigated the expression levels of several genes involved in metastasis and drug response in RENCA cells growing in ectopic (skin) and orthotopic (kidney) sites.
MATERIALS AND METHODS
Murine renal carcinoma cells were injected into kidney (orthotopic) and subcutis (ectopic) of syngeneic mice. Mice were treated with doxorubicin (DXR) (8 mg/kg) on days 8 and 15 after tumor cell implantation. Drug response was measured both in vivo and ex vivo by measuring tumor size and MTT assay. We also performed an in situ mRNA hybridization to estimate the expression levels of mdr (multidrug resistance), EGFR (epidermal growth factor receptor) and type IV collagenase.
RESULTS
RENCA cells growing in the kidney of syngeneic mice produced metastatic lesions in the lung (57% of mice), while the same cells growing in the subcutis did not. Tumors growing in the kidney were more resistant to DXR than tumors growing in the subcutis. MTT assays revealed that tumor cells derived from kidney were more resistant to DXR than those cells from subcutis. In situ hybridization analyses showed that transcripts of EGFR and type IV collagenase genes in kidney tumors were higher than those of subcutaneous tumors but mdr expression showed no difference between the two tumors.
CONCLUSION
These results demonstrate that the organ environment influences the drug responsive ness and the expression of EGFR and type IV collagenase genes in murine renal cell carcinoma cells.
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Matrix Metalloproteinase Expression and Ldegrees Calization in Human Prostate Adendegrees Carcinoma: An Immunohistdegrees Chemical Study of MMP-2, -9, -3, -7
Jung Weon Shim, Young Goo Lee, Tae Jin Lee, Jae Y Ro
J Korean Cancer Assoc. 2000;32(3):629-637.
AbstractAbstract PDF
No abstract available.
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Clinical Significance of Apoptosis and p53 Protein Expression in Stage IIB Squamous Cell Carcinoma of the Cervix Treated with Radiotherapy Alone
Eun Ji Chung, Gwi Eon Kim, Jinsil Seong, Woo Ick Yang, Young Tae Kim, Chang Ok Suh
J Korean Cancer Assoc. 2000;32(3):638-646.
AbstractAbstract PDF
PURPOSE
The purpose was to investigate the spontaneous apoptotic index (SAI) and p53 protein expression and to identify the role of SAI and p53 protein positivity.
MATERIALS AND METHODS
Forty six patients with squamous cell carcinoma of the cervix, FIGO stage IIB, treated with curative radiotherapy alone between 1990 and 1993 were included in this study. Definitive radiotherapy including external beam and high-dose-rate brachytherapy was given. Pretreatment paraffin-embedded biopsy specimens of those patients were scored for apoptosis and p53 protein expression using mouse mondegrees Clonal antibody (DO-7) by immuno staining. Clinicopathologic characteristics were also studied in relation to SAI and p53 protein expression, and as prognostic factors for clinical outcome.
RESULTS
SAI and p53 were not related to any clinical characteristics. The range of the SAI was 0.2~4.7% (median 1.1%, mean 1.5%). The rate of p53 protein expression was 65.2% (30/46). Patients whose tumors had high SAI and low p53 protein positivity had better treatment outcome than those with lower SAI. There was also a significant correlation between the SAI and p53 protein expression.
CONCLUSION
The pretreatment SAI and p53 oncoprotein expression are clinically useful in predicting the clinical outcome of FIGO stage IIB squamous cell carcinoma of the uterine cervix patients treated with definitive radiotherapy.
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Hematopoietic Recovery of Peripheral Blood Stem Cells Stored at 4degrees C
Seok Goo Cho, Eun Jee Oh, Jun Mo Lee, Hoon Kyo Kim, Kyung Shick Lee, Chun Choo Kim
J Korean Cancer Assoc. 2000;32(3):647-654.
AbstractAbstract PDF
PURPOSE
Cryopreservation has been the standard method of storing hematopoietic cells for the past 20 years, but this prdegrees Cedure is laborious and expensive. So, we evaluated the hematopoietic recovery of stored PBSCs at 4degrees C for a variable storage period MATERIALS AND METHODS: Eight leukapheresis products were kept unprdegrees Cessed at 4degrees C for 96 hours. To evaluate the effect of storage period on the hematopoietic recovery of PBSCs, assays for viability of mononuclear cells (MNCs), CFU-GM colony counts and CD34 cell counts were performed every 24 hours after PBSC collection. We tried to compare hematopoetic recovery of stored PBSCs at 4degrees C with that of cryopreserved PBSCs by using repeated measures ANOVA.
RESULTS
Viability of MNCs, CFU-GM colony counts and CD34 cell counts were monitored at 24 hour, 48 hour, 72 hour and 96 hour after PBSC collection. Data are expressed as percentage of baseline value and shown as mean s.d.; MNCs viability (96+/-2%, 94+/-2%, 92+/-2%, 88+/- 3%), CFU-GM colony counts (87+/-10%, 79+/-11%, 65+/-13%, 56+/-15%), and CD34 cell counts (93+/-13%, 93+/-12%, 88+/-14%, 85+/-19%). After storing PBSCs at 4degrees C for 96 hours, viability of MNCs and CFU-GM colony counts were significantly reduced (p<0.05) except CD34 cell concentration (p>0.05). Prdegrees Cedures of controlled-rate freezing and thawing resulted in a notable loss of viability (77+/-9%) and CFU-GM colony count (71+/-29%). CFU-GM colony counts of 72 hour-stored PBSCs at 4degrees C was similar to those of cryopreserved PBSCs.
CONCLUSION
If G-CSF mobilized PBSCs are stored at 4degrees C in less than 72 hours after collection, those hematopoietic recovery would be comparable to that of cryopreserved stem cells which are achieved by the rate-control freezer.
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Screening and Cloning of Genes Preferentially Expressed in Aminopterin-Treated Myeloma Cell Apoptosis
Hong Chan Lee, Sung Joo Kim, Sarah Yoon, Sung Joon Kwon, Yong Hoon Chung
J Korean Cancer Assoc. 2000;32(3):655-664.
AbstractAbstract
PURPOSE
When murine myeloma cells P3-X63-Ag8.653 (V653) of this model treated with amin opterin, an anticancer drug, they can't synthesize nucleic acid via de novo or salvage pathway and selectively eliminated due to apoptosis. This study was aimed to clone specific known and novel genes preferentially expressed in aminopteirn-treated tumor cell apoptosis.
MATERIALS AND METHODS
This study was aimed to clone specific known and novel genes pre ferentially expressed in aminopteirn-treated tumor cell apoptosis by using subtraction-PCR technique.
RESULTS
By using this technique 868 clones were obtained. Of these 427 clones were positive with insert DNA. By using cross-hybridization Southern blotting, final 101 clones were selected. All of these genes were sequenced and analyzed by using genebank DNA database. Total 101 clones of genes preferentially expressed in apoptotic tumor cells were classified into 10 groups, which included ribosomal proteins, nuclear proteins, mitdegrees Chondrial proteins, signal transductional proteins, retroviral proteins, cell surface receptor proteins, cell structural proteins, unclassified miscellaneous proteins, and novel genes. Especially, Unknown novel genes preferentially ex pressed in this apoptotic tumor cells included clone numbers S1-63, 1-1, 1-3, 1-16, 1-18, 1-20, 3-33, 3-41, 3-44, 3-48, 3-55, 3-60, 6-17, 6-25, 8-12, 50-7, 50-23, and 100-35.
CONCLUSION
It seemed that known and unknown novel genes cloned in this study would con tribute to the future studies regarding apoptosis of tumor cells and cancer treatment therepy.
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Cancer Res Treat : Cancer Research and Treatment
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