摘要
目的探讨雷氏大疣蛛M acrothele raven i毒素对人肝癌细胞SMM C-7721增殖及细胞周期的抑制作用,进一步探讨其作用的分子机制。方法采用M TT法测定雷氏大疣蛛毒素对SMM C-7721细胞增殖作用的影响;采用[3H]-T dR掺入法检测雷氏大疣蛛毒素作用前后SMM C-7721细胞DNA合成的变化;采用流式细胞术(FCM)探讨雷氏大疣蛛毒素对SMM C-7721细胞凋亡率和细胞周期的影响;采用W estern B lot方法研究雷氏大疣蛛毒素对细胞周期相关c-m yc蛋白表达的影响。结果M TT方法表明雷氏大疣蛛毒素对SMM C-7721细胞增殖有较强的抑制作用(P<0.05),时效和量效关系良好。雷氏大疣蛛毒素可以抑制SMM C-7721细胞DNA的合成。流式细胞仪检测结果发现雷氏大疣蛛毒素作用后SMM C-7721细胞凋亡率增加,细胞周期阻滞在G2/M期。W estern B lot方法进一步检测到雷氏大疣蛛毒素作用SMM C-7721细胞72 h后,c-m yc蛋白表达减弱。结论雷氏大疣蛛毒素可以抑制人肝癌细胞SMM C-7721的增殖和DNA的合成,其机制可能是诱导细胞凋亡,使细胞周期相关c-m yc蛋白表达减弱,导致细胞周期的变化。
Objective To study the effect of spider venom from Macrothele raveni on proliferation and cell cycles of human hepatocellular carcinoma cell line SMMC-7721 and the molecular mechanism of the effect. Methods Proliferation of SMMC-7721 cells was determined by MTT assay. DNA synthesis of SMMC-7721 cell pre- and post-treatment with spider venom from M. raveni was tested by [^3H] TdR assay. The induction of apoptosis and the change of cell cycle in SMMC-7721 cells treated with spider venom from M. raveni were investigated by Flow Cytometry. The effect of spider venom from M. raveni on expression of c-myc protein in SMMC-7721 cells was studied by Western Blot. Results MTT assay showed that the proliferation of SMMC-7721 cells in vitro was inhibited by spider venom from M. raveni （P〈0.05） in time-dependent and concentration-dependent manners. The DNA synthesis of SMMC-7721 cells was inhibited by spider venom from M. raveni. The apoptosis and inhibition of SMMC 7721 cells in the G2/M phase were induced after treated with the spider venom from M. raveni. Expression of c-myc protein in SMMC 7721 cells treated with spider venom from M. raveni for 72 h was obviously down- regulated. Conclusion Spider venom from M. raveni could inhibit the proliferation and DNA synthesis of SMMC 7721 cells. The potential mechanisms are to induce cell apoptosis, activate the lower expression of c-myc protein and change the cell cycle of SMMC-7721 cells.
出处
《中草药》
CAS
CSCD
北大核心
2006年第1期74-77,共4页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金项目(30371753)
河北省教育厅科研项目(2004110)
河北师范大学科研基金资助
