姜黄素协同二甲双胍抑制人宫颈癌细胞增殖及作用机制

Mechanism of Curcumin Combined with Metformin Inhibiting Proliferation of Human Cervical Cancer Cells

目的探究姜黄素协同二甲双胍抑制人宫颈癌细胞增殖及作用的机制.方法体外培养人宫颈癌HeLa细胞,采用MTT法检测不同浓度的姜黄素及二甲双胍对HeLa细胞抑制率并绘制12、24及48 h细胞抑制率曲线;将各组HeLa细胞分为空白对照组、姜黄素组、二甲双胍组和联合组;姜黄素组使用25μmol/L的姜黄素处理,二甲双胍组使用1 mmol/L的二甲双胍处理,联合组使用25μmol/L的姜黄素+1 mmol/L的二甲双胍处理;采用克隆形成实验检测各组HeLa细胞的增殖情况;使用WB检测增殖相关蛋白Ki67和PCNA表达情况;使用流式细胞仪检测各组细胞不同细胞周期的细胞占比.结果MTT法检测发现在一定浓度范围内,姜黄素及二甲双胍对HeLa细胞抑制率呈浓度依赖性,姜黄素对宫颈癌细胞的半数致死浓度(IC50)为25.97μmol/L,二甲双胍对宫颈癌细胞的IC50为0.985 mmol/L;相比空白对照组,姜黄素组和二甲双胍组宫颈癌HeLa细胞克隆形成率、Ki67蛋白相对表达量、PCNA蛋白相对表达量和S期细胞比例显著降低(P<0.05),G0/G1期、G2/M期细胞比例和细胞凋亡率显著升高(P<0.05);相比姜黄素组和二甲双胍组,联合组宫颈癌HeLa细胞克隆形成率、Ki67蛋白相对表达量、PCNA蛋白相对表达量和S期细胞比例显著降低(P<0.05),G0/G1期、G2/M期细胞比例和细胞凋亡率显著升高(P<0.05).结论姜黄素协同二甲双胍可以抑制人宫颈癌细胞增殖,其作用机制可能与抑制增殖相关蛋白表达并将宫颈癌HeLa细胞阻滞于G0/G1期.

Objective This study explored the mechanism of curcumin combined with metformin inhibiting proliferation of human cervical cancer(CC)cells.Methods Human CC Hela cells were culturedin vitro.The inhibition of curcumin and metformin of different concentrations for Hela cells was detected by MITT.The curves of 12 h,24 h and 48 h cell inhibition rates were.drawn.Hela cells were divided into blank control group,curcumin group,metformin group and combination group.The curcumin group was treated with 25μumol/L curcumin,metformin group was treated with 1 mmol/L metformin,and combination group was treated with 25μmol/L curcumin in combination with 1 mmol/L metformin.The proliferation of Hela cells in each group was detected by clone formation assay.The expression of proliferation-related proteins(Ki67,PCNA)was detected by Western blotting.The proportions of cells in different cell cycles in each group were detected by flow cytometry.Results MTT showed that,within certain concentration range,inhibition rates of curcumin and metformin were concentration-dependent with with Hela cells.The half-inhibitory concentrations.(ICs)of curcumin and metformin for cervical cancer cells were 25.97μmol/L and 0.985 mmol/L,respectively.Compared with blank control group,clonal formation rate of CC Hela cells,relative expression levels of Ki67 and PCNA proteins,and cells proportions in S phase were significantly decreased in curcumin group and metformin group(P<0.05),while cells proportions in G0/G1 phase and G2/M stage,and apoptosis rate were significantly increased(P<0.05).Compared with curcumin group and metformin group,clonal formation rate of CC Hela cels,relative expression of Ki67 and PCNA proteins,and cells proportions in S phase were significantly decreased in combination group(P<0.05),while cells proportions in G0/G1 and G2/M phase,and apoptosis rate were significantly increased(P<0.05).Conclusion Curcumin combined with metformin can inhibit proliferation of human CC cells.The mechanism might be related to its ability to inhibit the expression of proliferation-related proteins and to arrest CC Hela cells at G0/G1 phase.