藤梨根乙酸乙酯制剂诱导食管癌细胞凋亡机制的观察

目的：探讨藤梨根（Actinidia arguta）乙酸乙酯制剂促进人食管癌Eca-109细胞凋亡的机制。方法采用MTT比色法，检测藤梨根乙酸乙酯药液在不同浓度（1μg/ml、10μg/ml、100μg/ml等）及不同时间（24h、48h、72h等）对Eca-109细胞生长抑制作用；采用TUNEL法检测其对癌细胞生长的诱导凋亡效应；采用免疫组化SP法，检测凋亡相关蛋白Bax 、Bcl-2及caspase的表达。结果藤梨根乙酸乙酯药液对人食管癌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%。藤梨根制剂对瘤细胞有明显的凋亡效应，而在对照组，未见有明显凋亡现象。瘤细胞作用24h、48h、72h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，72h后最明显，差异有统计学意义（P〈0.01）。藤梨根药液促进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。

{Abstract}： Objective To study the mechanism of Actinidia preparations on the apoptosis induction in human carcinoma of esophagus Eca-109 cells. Methods MTT colonmetric assay was used to examine the growth inhibition of concentration-effect（1μg/ml、10μg/ml、100μg/ml etc）and time-effect（24h、48h、72h etc）of extracts from Actinidia arguta by ethyl acetate on Eca-109 cells,TUNEL test were performed to observe the apoptosis induced by the extracts and the level of expression of Bax、Bcl-2 and caspase in tumor cells were measured by immunohistochemical method（SP technique）. Results The inhibitory effect of the extracts from Actinidia arguta by ethyl acetate on Eca-109 cells increased step by step to concentration and time,the highest rate of inhibition was 87.2%. The extracts can significantly induced apoptosis of Eca-109 cells,but in control group,apoptosis was not observed. Compared with control group,the level of expression Bax in tumor cells increased（P〈0.01）,but the level of expression of Bcl-2 decreased after 24h、48h and 72h（P〈0.01）,particularly after 72h. The expressions of caspase-3 and caspase-9 were significantly enhanced by Actinidia arguta,but Little effect on caspase-8. Conclusions Actinidia arguta may induces apoptotic Eca-109 cells death through suppressing bcl-2 and then activating caspase-9 and caspase-3.