藤梨根诱导人食管癌Eca-109细胞凋亡的调节机制

Regulatory Mechanisms of Apoptosis of Actinidia Arguta on Human Carcinoma of Esophagus Eca-109 Cells

目的研究藤梨根(Actinidia arguta)抑制人食管癌Eca-109细胞增殖和诱导凋亡的机制。方法采用MTT比色法,检测藤梨根正丁醇药液在不同浓度(1μg/ml、10μg/ml、100μg/ml等)及不同时间(24 h、48 h、72 h等)对Eca-109细胞生长抑制作用;采用免疫组化SP法,检测凋亡相关蛋白Bax、Bcl-2及Caspase的表达。结果藤梨根正丁醇药液对人食管癌Eca-109细胞的生长抑制作用,随着药物浓度的升高和作用时间的延长而增强,其生长抑制率可达87.2%。瘤细胞作用24 h﹑48 h﹑72 h后分别与对照组比较,用药组Bax蛋白表达明显增强(P<0.01);同时伴有Bcl-2表达减弱7,2 h后最明显,差异有显著意义(P<0.01)。藤梨根正丁醇药液促进Caspase-3、Caspase-9表达明显增强,而对Caspase-8表达影响较小。结论藤梨根可通过下调Bcl-2表达,激活Caspase-9、Caspase-3诱导Eca-109细胞凋亡。

Objective To study the mechanism of Actinidia arguta on the proliferation and apoptosis induction in human carcinoma of esophagus Eca-109 cells.Methods MTT colonmetric assay was used to examine the growth inhibition of concentration-effect（1 μg/ml、10 μg/ml、100 μg/ml） and time-effect（24 h﹑48 h﹑72 h）of extracts from Actinidia arguta by n-butyl alcohol on Eca-109 cells,the level of expression Bax、Bcl-2 and Caspase in tumor cells were measured by immunohistochemical method（SP technique）.Results The inhibitory effect of the extracts on Eca-109 cells increased step by step to concentration and time,the highest rate of inhibition was 87.2%.Compared with control group,the level of expression Bax in tumor cells increased（P0.01）,but the level of expression of Bcl-2 decreased after 24 h﹑48 h and 72 h（P0.01）,particularly after 72h.The expressions of Caspase-3 and Caspase-9 significantly enhanced by Actinidia arguta,but Little effect on Caspase-8.Conclusion Actinidia arguta may induces apoptotic Eca-109 cells death through suppressing bcl-2 and then activating Caspase-9 and Caspase-3.