姜黄素联合顺铂调控PI3K/Akt信号通路对卵巢癌耐药细胞增殖、迁移和凋亡的影响

Effect of curcumin combined with cisplatin on PI3K/Akt signaling pathway on proliferation,migration and apoptosis of drug-resistant ovarian cancer cells

目的:讨论姜黄素联合顺铂对卵巢癌耐药细胞增殖、迁移及凋亡能力的作用,并分析其与PI3K/Akt通路的关系。方法:首先用不同浓度的姜黄素及顺铂单独处理细胞株,根据各浓度细胞的存活率确定35µmol/L姜黄素、4.0 mg/L顺铂疗效最好,用于后续实验。将卵巢癌耐药细胞分为空白对照组、姜黄素单独处理组(35µmol/L)、顺铂单独处理组(4.0 mg/L)、姜黄素+顺铂联合处理组(35µmol/L+4.0 mg/L),采用MTT法、Transwell小室、流式细胞术检测四组细胞的生长抑制率、迁移细胞数及细胞凋亡率;Western blot检测四组细胞中p-PI3K及p-Akt蛋白表达水平;PCR检测四组细胞中PI3K、Akt、Ki67、Bcl-2、Bax mRNA表达水平。结果:与空白对照组比较,其余三组细胞生长抑制率及细胞凋亡率均显著升高,迁移细胞数显著减少(P<0.05),且姜黄素+顺铂联合组细胞生长抑制率及细胞凋亡率显著高于姜黄素组、顺铂组,迁移细胞数则低于姜黄素组、顺铂组(P<0.05);与空白对照组比较,姜黄素处理组及姜黄素+顺铂联合组细胞中p-PI3K、p-Akt蛋白及PI3K、Akt、Ki67、Bcl-2 mRNA表达水平均显著降低、Bax mRNA显著升高(P<0.05),且联合组细胞表达量明显低于姜黄素处理组(P<0.05),而顺铂单独处理组细胞中PI3K、p-Akt蛋白及PI3K、Akt、Ki67、Bcl-2、Bax mRNA表达水平无明显变化(P>0.05)。结论:姜黄素联合顺铂能够提高卵巢癌耐药细胞的敏感性,抑制细胞的增殖及迁移,同时诱导细胞凋亡,这一过程可能通过下调PI3K/Akt通路的表达实现。

Objective:To discuss the effects of curcumin combined with cisplatin on proliferation,migration and apoptosis of drug-resistant ovarian cancer cells,and to analyze its relationship with PI3K/Akt pathway.Methods:Firstly,the cell lines were treated with different concentrations of curcumin and cisplatin separately,and according to the survival rate of cells of each concentration,it was determined that 35µmol/L curcumin and 4.0 mg/L cisplatin had the best effect,which was used in subsequent experiments.Drugresistant ovarian cancer cells were divided into blank control group,curcumin single treatment group(35µmol/L),cisplatin single treatment group(4.0 mg/L),curcumin+cisplatin combined treatment group(35µmol/L+4.0 mg/L),MTT method,Transwell chamber,flow cytometry were used to detect growth inhibition rate,number of migrating cells and apoptosis rate of four groups of cells;Western blot was used to detect expression levels of p-PI3K and p-Akt in four groups of cells;expression levels of PI3K,Akt,Ki67,Bcl-2 and Bax mRNA in four groups of cells were detected by PCR.Results:Compared with blank control group,cell growth inhibition rate and apoptosis rate of other three groups were significantly increased,while the number of migrating cells was significantly decreased(P<0.05),and cell growth inhibition rate and apoptosis rate of curcumin+cisplatin combination group were significantly higher than that of curcumin group and cisplatin group,while the number of migrating cells was lower than that of curcumin group and cisplatin group(P<0.05);compared with blank control group,expression levels of p-PI3K,p-Akt protein and PI3K,Akt,Ki67 and Bcl-2 mRNA in cells of curcumin treatment group and curcumin+cisplatin combination group were significantly decreased,while Bax mRNA was signifi-cantly increased(P<0.05),and cell expression of combination group was significantly lower in curcumin treatment group(P<0.05),while PI3K,p-Akt protein and PI3K,Akt,Ki67,Bcl-2,Bax mRNA expression levels in cells of cisplatin treatment group did not change significantly(P>0.05).Conclusion:Curcumin combined with cisplatin can increase sensitivity of drug-resistant ovarian cancer cells,inhibit cell proliferation and migration,and induce apoptosis at the same time.This process may be achieved by downregulating the expression of PI3K/Akt pathway.