表没食子儿茶素没食子酸酯测定及其对胃癌细胞增殖抑制作用探究.doc

表没食子儿茶素没食子酸酯测定及其对胃癌细胞增殖抑制作用探究 　 作者：张凤，安利国，李福荣，赵晓民，文今福，杨桂文 【摘要】 目的分析表没食子儿茶素没食子酸酯(EGCG)在绿茶提取物中的含量，探讨EGCG对人胃癌细胞BGC823增殖和凋亡的影响，为阐明EGCG抗肿瘤作用机制提供实验基础。方法反相高效液相色谱法(RP-HPLC)测定绿茶提取物中EGCG的含量。MTT法检测EGCG对体外培养的人胃癌细胞BGC823增殖的影响;DNA ladder法检测EGCG处理各组DNA梯度的变化。所有数据均采用SPSS12.0进行统计分析，均数比较采用t-test。结果通过绘制标准曲线确定绿茶提取物中EGCG 的含量为9%。MTT法检测结果显示EGCG能够呈现剂量(5×10-3,1×10-2,2×10-2,4×10-2)g/L依赖性地抑制人胃癌细胞BGC823的增殖;DNA ladder法显示EGCG处理组细胞DNA呈现梯状条带，而对照组则没有这种带纹。结论绿茶提取物中EGCG的含量为9%;EGCG能够通过诱导细胞凋亡而抑制人胃癌细胞BGC823的生长和增殖。 【关键词】 反相高效液相色谱; 表没食子儿茶素没食子酸酯; 胃癌细胞; 细胞凋亡 　　Abstract：ObjectiveTo analyze the content of Epigallocatechin gallate(EGCG)in green tea extraction and to investigate the effect of EGCG on the proliferation and apoptosis of human gastric cancer cells BGC823 cultured in vitro,which provide experimental basis for clarifying the mechanism of antitumor effect of EGCG.MethodsIn the condition of mobile phase double distilled water / acetonitrile / ethyl acetate (86∶12∶2 v / v / v), a flow rate of 1.0 ml / min, RP-HPLC Chromatogram was used to study EGCG in green tea. Human gastric cancer cells BGC823 cultured in vitro were treated with different concentrations of EGCG, and then the absorbencyon at 490 nm of the cells in each group was examined by MTT method. DNA ladder assay was used to detect the apoptosis of BGC823.Statistical analysis was performed by SPSS12.0 and t test was used to compare the mean value between the different groups.ResultsThere was 9% EGCG in green tea extraction calculating by drawing standard cure. After treatment with different concentrations of EGCG, as measured by MTT assay ,cell viability decreased in a dose- (5×10-3,1×10-2,2×10-2,4×10-2g/L) dependent manner(showed as figure 2). DNA ladder assay showed that DNA of the cells treated with EGCG showed the characteristics of apoptosis-DNA ladder,while the control group did not have such strip.ConclusionThere was 9% EGCG in green tea.EGCG can inhibit the growth and proliferation of gastric cancer cells BGC823 by inducing its apoptosis