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Original Articles
- 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.
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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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- 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.
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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.
- 3,103 View
- 31 Download
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