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作 者:曹玲[1] 钱亚云[1] 刘延庆[1] 朱耀东[1] 杨林[1] 陈珏[1] 顾昊[1] 白云飞[1] 金凤[1]
出 处:《中国实验方剂学杂志》2015年第14期97-101,共5页Chinese Journal of Experimental Traditional Medical Formulae
基 金:国家自然科学基金项目(81274141;81403232);江苏省自然科学基金项目(BK2012686)
摘 要: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 effects of Celastrus orbiculatus extract (COE) on apoptosis in human gastric carcinoma MGC-803 cells in which the tumor suppressor gene, maspin, is overexpressed. Method: The growth rates of maspin-overexpressed human gastric carcinoma MGC-803 cells treated with different concentrations of COE (20, 40, 80, 160, 320 mg·L-1) for 24 and 48 h were measured by MTT assay. The apoptosis of MGC-803 ceils treated with different concentrations of COE (0, 20, 40, 80 mg ·L-1 ) and 5-FU (32 mg ·L -1 ) for 24 h was examined according to the Tunel Apoptosis Assay Kit. The protein expressions of maspin, Bcl-2, Bcl-2LI2 and Bax were detected by using Western blot. Result: COE (20, 40, 80, 160, 320 mg ·L-1 ) suppressed the proliferation of maspin-overexpressed human gastric cancer MGC-803 cells in a concentration-and time-dependent manner. After being treated with COE (20, 40, 80 mg ·L-1 ) for 24 h, reduction of cell number, morphology being round, cytoplasmic concentration and apoptotic bodies were observed gradually with the growing of the concentration. Tunel assay showed that COE (20, 40, 80 mg ·L-1 ) could promote apoptosis of cells. Western blot showed that COE (20, 40, 80, 160 mg ·L-1) increased the expression of maspin and decreased expressions of Bcl-2 and Bcl-2L12 in a concertration-dependent manner. Conclusion: COE could induce the apoptosis of human gastric cancer MGC-803 cells in which maspin gene is overexpressed, the mechanisms may be related to regulating the expressions of Bcl-2, Bcl-2L12 and maspin.
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