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目的:利用高表达抑癌基因maspin的人胃癌MGC-803细胞探讨南蛇藤提取物对其凋亡的影响。方法:以人胃癌MGC-803细胞为研究对象,不同质量浓度20,40,80,160,320 mg·L-1南蛇藤提取物作用于MGC-803细胞24,48 h后,MTT法检测南蛇藤提取物对高表达maspin的人胃癌MGC-803细胞增殖的影响;以不同质量浓度COE组(0,20,40,80 mg·L-1)及32 mg·L-1的5-FU阳性对照处理24 h后,TUNEL法检测南蛇藤提取物对细胞凋亡的影响,Western blot检测对maspin,Bcl-2,Bcl-2L12及Bax蛋白表达的影响。结果:南蛇藤提取物(20,40,80,160,320 mg·L-1)能明显抑制高表达maspin的人胃癌MGC-803细胞的增殖,呈现时间及浓度依赖性;与空白组比较,不同质量浓度的南蛇藤提取物(20,40,80 mg·L-1)作用24 h后,镜下显示,随着药物浓度升高,细胞数目减少,形态变圆,胞浆浓缩,出现凋亡小体,TUNEL结果表明南蛇藤提取物(20,40,80 mg·L-1)能促进细胞凋亡,Western blot结果显示南蛇藤提取物(20,40,80,160 mg·L-1)可呈浓度依赖性地增加抑癌基因maspin蛋白的表达,降低Bcl-2,Bcl-2L12的表达,均具有明显的统计学差异(P<0.05,P<0.01)。结论:南蛇藤提取物能够抑制高表达maspin的人胃癌MGC-803细胞的增殖,促进其凋亡,其机制可能与南蛇藤提取物促进抑癌基因maspin的表达以及抑制Bcl-2,Bcl-2L12的表达有关。
OBJECTIVE: To investigate the effect of extract of Celastrus orbiculatus on its apoptosis using human gastric cancer MGC-803 cell line with high expression of the tumor suppressor gene maspin. Methods: Human gastric cancer MGC-803 cells were treated with 20, 40, 80, 160 and 320 mg · L-1 of Celastrus orbiculatus extracts at different concentrations for 24 and 48 h. MTT assay On the proliferation of human gastric cancer cell line MGC-803 with high expression of maspin. The cells were treated with different concentrations of COE (0, 20, 40, and 80 mg · L -1) and 5-FU positive control Twenty-four hours later, the effects of Celastrus orbiculatus extract on apoptosis were detected by TUNEL method. The protein expressions of maspin, Bcl-2, Bcl-2L12 and Bax were detected by Western blot. Results: The extract of Celastrus orbiculatus (20,40,80,160,320 mg · L-1) could significantly inhibit the proliferation of human gastric cancer MGC-803 cells with high maspin expression in a time and concentration dependent manner. Compared with the blank group, (20, 40, 80 mg · L-1) for 24 h, microscopic examination showed that with the increase of drug concentration, the number of cells decreased, the morphology became round, and the cytoplasm was concentrated and apoptosis was small The results of TUNEL showed that Celastrus orbiculatus extract (20, 40, 80 mg · L-1) could promote cell apoptosis. Western blot results showed that Celastrus orbiculatus extract (20, 40, 80 and 160 mg · L -1) There was a statistically significant difference (P <0.05, P <0.01) in increasing the expression of the tumor suppressor gene maspin protein and decreasing the expression of Bcl-2 and Bcl-2L12 in a concentration-dependent manner. CONCLUSION: The extract of Celastrus orbiculatus can inhibit the proliferation and promote the apoptosis of human gastric cancer cell line MGC-803 with high expression of maspin, which may be related to the effect of Celastrus orbiculatus extract on promoting the expression of the tumor suppressor gene maspin and inhibiting the expression of Bcl-2, Bcl- -2L12 expression.