扶正解毒祛瘀方联合奥沙利铂对人结肠癌HT-29细胞增殖与凋亡的影响及机制研究
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篇名: 扶正解毒祛瘀方联合奥沙利铂对人结肠癌HT-29细胞增殖与凋亡的影响及机制研究
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摘要: 目的:研究扶正解毒祛瘀方(简称“扶正方”)联合奥沙利铂(L-OHP)对人结肠癌HT-29细胞增殖与凋亡的影响及机制。方法:将HT-29细胞分为空白对照组(不加药物)、扶正方组(1 000 mg/L)、L-OHP组(31.25 mg/L)和联合用药组(1 000 mg/L扶正方+31.25 mg/L L-OHP)。加入相应药物作用48 h后,采用MTT法检测细胞增殖情况,倒置显微镜下观察细胞形态的改变,流式细胞术检测细胞周期和凋亡率,实时荧光定量聚合酶链式反应(qRT-PCR)法检测细胞中促凋亡基因Bax、抑凋亡基因Bcl-2 mRNA的表达,Western blot法检测细胞中Bax、Bcl-2蛋白的表达。结果:与空白对照组比较,L-OHP组和联合用药组细胞增殖均受到抑制、S期和G2/M期细胞比例升高、G0/G1期细胞比例降低(P<0.05),L-OHP组、扶正方组和联合用药组细胞凋亡率升高、细胞中Bax mRNA及蛋白的表达上调、细胞中Bcl-2 mRNA的表达下调(P<0.05),且联合用组变化较两药单用组更明显(P<0.05)。结论:扶正方与L-OHP联合应用可抑制HT-29细胞的增殖、促进细胞凋亡,且作用优于两药单用;其机制可能与上调细胞中Bax 基因与蛋白表达、下调细胞中Bcl-2 基因表达有关。
ABSTRACT: OBJECTIVE: To study the effect and its mechanism of Fuzheng Jiedu Quyu formula (short for “Fuzheng formula”) combined with oxaliplatin (L-OHP) on human colon cancer HT-29 cell proliferation and apoptosis. METHODS: HT-29 cells were divided into blank control group (without drugs), Fuzheng formula group (1 000 mg/L), L-OHP group (31.25 mg/L) and combination group (1 000 mg/L Fuzheng formula+31.25 mg/L L-OHP). After cultured with corresponding drug for 48 h, MTT method was used to detect the cell proliferation; the changes of cellular morphology were observed by invert microscope; flow cytometry was used to detect the cell cycle and apoptosis rate. Proapoptotic gene Bax, apoptotic gene Bcl-2 mRNA expressions were determined by real-time fluorescence quantitative polymerase chain reaction method; Bax, Bcl-2 protein expressions were assayed by Western blot. RESULTS: Compared with blank control group, cell proliferation was inhibited in L-OHP group and combination group; cell proportion was increased in S stage, G2/M stage and decreased in G0/G1 stage (P<0.05). Cell apoptosis rate in L-OHP group, Fuzheng formula group and combination group was increased; Bax mRNA and protein expression were up-regulated, Bcl-2 mRNA expression was downregulated (P<0.05); and combination group changed more obviously than the single drug groups (P<0.05). CONCLUSIONS: Fuzheng formula combined with L-OHP can inhibit HT-29 cell proliferation and promote its apoptosis, showing better effects than either of the two drugs alone. The mechanism may be associated with up-regulation of Bax gene and protein expressions and down-regulation of Bcl-2 gene expressions in cells.
期刊: 2017年第28卷第19期
作者: 赵秀梅,周冰,张桂贤,柴仲秋,刘晓芸,刘洪斌
AUTHORS: ZHAO Xiumei,ZHOU Bing,ZHANG Guixian,CHAI Zhongqiu,LIU Xiaoyun,LIU Hongbin
关键字: 扶正解毒祛瘀方;奥沙利铂;人结肠癌HT-29细胞;增殖;凋亡;体外
KEYWORDS: Fuzheng Jiedu Quyu formula; Oxaliplatin; Human colon cancer HT-29 cell; Proliferation; Apoptosis; in vitro
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