南蛇藤提取物诱导SGC-7901胃癌细胞凋亡及其机制

Apoptosis of Gastric Cancer SGC-7901 Cells Induced by Celastrus orbiculatus Thunb Extract and Mechanism of Induction

目的观察南蛇藤乙酸乙酯提取物和正丁醇提取物在体外诱导SGC-7901胃癌细胞凋亡的作用,并初步探讨其分子机制。方法制备南蛇藤乙酸乙酯和正丁醇提取物,采用MTT法检测二者对SGC-7901胃癌细胞增殖的影响;应用FITC-AnnexinⅤ及碘化丙锭(PI)双标法,通过流式细胞仪(FCM)观察SGC-7901细胞的凋亡率及P53蛋白的表达。结果不同浓度的南蛇藤乙酸乙酯和正丁醇提取物(15、30、60、120μg/ml)对SGC-7901细胞的增殖均有一定的抑制作用,且呈剂量依赖性。两种南蛇藤提取物(60、120、240、480μg/ml)可剂量依赖性地诱导肿瘤细胞凋亡,且随着浓度的升高,SGC-7901细胞P53蛋白的表达也增加,浓度为240和480μg/ml的提取物组与对照组相比,差异有统计学意义。结论南蛇藤乙酸乙酯提取物和正丁醇提取物在体外可诱导SGC-7901细胞凋亡,上调细胞中P53基因的表达可能是其抗肿瘤作用的分子机制之一。

Objective To observe the apoptosis of gastric cancer SGC-7901 cells induced in vitro by extracts from Celastrus orbiculatus thunb with ethyl acetate and n-butanol and investigate the relevant molecular mechanism. Methods Celastrus orbiculatus thunb was extracted with ethyl acetate and n-butanol, and the effect of extracts on proliferation of SGC-7901 cells were determined by MTT method. The apoptosis of SGC-7901 cells and expression of P53 protein were determined by double labeled technique with FITC-Annexin V and PI and flow cytometry. Results Celastrus orbiculatus thunb extracts showed dosedependent inhibitory effect on the proliferation of SGC-7901 cells. Both extracts with ethyl acetate and n-butanol induced the apoptosis of SGC-7901 cells in dose-dependent modes. The expression level of P53 protein in SGC-7901 cells increased with the increasing concentrations of extracts. The fluorescence indexes （FIs）of P53 protein in SGC-7901 cells treated with either extract at concentrations of 240 and 480 μg / ml were significantly different from those of negative control. Conclusion Both the extracts from Celastrus orbiculatus thunb with ethyl acetate and n-butanol induced the apoptosis of SGC-7901 cells in vitro by a potential mechanism of up-regulating the expression of P53 gene.