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雷氏大疣蛛毒素对人肝癌细胞SMMC-7721细胞增殖及细胞周期的影响

Effect of spider venom from Macrothele raveni on proliferation and cell cycle of human hepatocellular carcinoma cell line SMMC-7721

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【作者】 孙捷高莉沈永青单保恩

【Author】 SUN Jie~1,GAO Li~2,SHEN Yong-qing~3,SHAN Bao-en~4(1.Science and Technology Company,Hebei Medical University,Shijiazhuang 050017,China;2.College of Life Science,Hebei Normal University,Shijiazhuang 050016,China;3.Department of Oncology, People’s Hospitalof Hebei Province,Shijiazhuang 050051,China;4.Research Centre,The Fourth Hospitalof Hebei Medical University,Shijiazhuang 050011,China)

【机构】 河北医科大学科技总公司河北师范大学生命科学学院河北省人民医院肿瘤科河北医科大学第四医院科研中心 河北石家庄050017河北石家庄050016河北石家庄050051河北石家庄050011

【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。

【Abstract】 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 -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.

【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
  • 【文献出处】 中草药 ,Chinese Traditional and Herbal Drugs , 编辑部邮箱 ,2006年01期
  • 【分类号】R735.7
  • 【被引频次】6
  • 【下载频次】156
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