摘要
[目的]研究绿茶多酚(EGCG)诱导肝癌细胞Hep-G2凋亡过程中半胱天冬酶-3(Caspase-3)蛋白活性与mRNA的变化。[方法]100 mg/L的EGCG处理Hep-G2细胞48 h,DAPI染色观察细胞形态学,用MTT检测EGCG对Hep-G2细胞生长的影响,流式细胞技术Annexin V-FITC PI法检测不同浓度EGCG诱导细胞的凋亡率,分光光度法检测Caspase-3的活性,用半定量RT-PCR法检测Caspase-3的mRNA变化。[结果]100 mg/L的EGCG作用Hep-G2细胞48 h后,荧光显微镜观察到细胞出现核碎裂和凋亡小体;MTT法检测到EGCG对肝癌细胞Hep-G2的生长均有显著抑制作用,呈浓度依赖性;Annexin V-FITC PI法检测到不同浓度的EGCG作用与肝癌细胞Hep-G2后,凋亡率较对照组明显升高,并随诱导浓度的增加而增加;不同浓度的EGCG处理组和不同时间EGCG处理组的Caspase-3活性和mRNA表达量均比对照组明显升高。[结论]Caspase-3蛋白活性与mRNA上调是EGCG诱导肝癌细胞凋亡的的重要原因和机制。
[ Objective ] This paper was conducted to analyze the activity of Caspase-3 and mRNA translation in the EGCG-induced apoptotic. [ Method] Hep-G2 cells were incubated with EGCG 100 mg/L for 48 h. The cellular morphological analysis was conducted by DAlai pigmentation. The infection of EGCG on the growth of Hep-G2 was tested by MTT. The cellular apoptotic rate was analyzed at different dose by a flow cytometry（Annexin V-FITC PI）. Activity of caspase-3 was measured using a colorimetric method. The mRNA level was semi-quantified by a reverse transcriptase-polymerase chain reaction technique. [ Result] After treated with EGCG 100 mg/L for 48 h, the Hep-G2 cells declined its shape with regard to nuclear fragmentation and apoptotic body by fluorescence microscope. EGCG inhibited proliferation of Hep-G2 cells in dose by MTT assay. Compared with the control, the apoptosis ratio of Hep-G2 cells incubated with EGCG increased by dose. The activity of caspase-3 and the mRNA level of the EGCG-treated Hep-G2 cells at different times and different doses were significantly higher than the those of control （P 〈 0.01 ）. [ Conclusion] EGCG led to the apoptosis of Hep-G2 cells by up-regulating activity of caspase-3 and mRNA level.
出处
《安徽农业科学》
CAS
北大核心
2008年第23期9907-9909,9997,共4页
Journal of Anhui Agricultural Sciences
基金
河南省自然科学基金项目(0624410029)
