鼠尾草酸联合他莫昔芬通过激活Caspase-3信号通路诱导乳腺癌细胞凋亡

摘要

目的 探讨鼠尾草酸联合他莫昔芬对乳腺癌细胞的影响及作用机制.方法 将乳腺癌细胞株MCF-7、T47D分为4组,对照组加入PBS,CA组加入鼠尾草酸,TAM组加入他莫昔芬,CA+TAM组加入鼠尾草酸和他莫昔芬.MTT增殖实验检测细胞增殖能力,集落形成实验检测细胞集落形成能力,Transwell小室实验检测细胞侵袭能力,细胞划痕实验检测细胞迁移能力,免疫印迹法检测各组细胞Caspase-8、Caspase-3、PARP蛋白表达.结果 在0~20 μmol/L药物浓度时,CA、TAM、CA+TAM呈剂量依赖性抑制乳腺癌细胞增殖,但药物浓度为20 μmol/L与50 μmol/L对细胞增殖影响差异无统计学意义(均P>0.05).CA+TAM组增殖抑制率明显高于CA组和TAM组(均P<0.05).MCF-7和T47D细胞中,CA组、TAM组、CA+TAM组集落形成率、侵袭细胞数、细胞迁移率显著低于对照组(均P<0.05);CA+TAM组集落形成率、侵袭细胞数、细胞迁移率显著低于CA组和TAM组(均P<0.05),CA组和TAM组集落形成率、侵袭细胞数、细胞迁移率比较差异均无统计学意义(均P>0.05).在2种乳腺癌细胞株中,对照组、CA组、TAM组、CA+TAM组cleaved Caspase-8蛋白表达差异无统计学意义(均P>0.05),CA组、TAM组、CA+TAM组cleaved Caspase-3、cleaved PARP蛋白表达显著高于对照组(均P<0.05),CA+TAM组cleaved Caspase-3、cleaved PARP蛋白表达显著高于CA组和TAM组(均P<0.05),CA组和TAM组cleaved Caspase-3、cleaved PARP蛋白表达差异无统计学意义(均P>0.05).结论 鼠尾草酸联合他莫昔芬能够抑制乳腺癌细胞的增殖、迁移和侵袭,其作用机制可能与激活Caspase-3/PARP信号通路有关.%Objective To explore the effect of carnosic acid combined with tamoxifen on breast cancer cells and the mechanism.Methods Breast cancer cell lines MCF-7,T47D were divided into 4 groups:PBS was given to control group,carnosic acid(CA)was added to CA group,tamoxifen(TAM)was added to TAM group,and CA+TAM group was treated with carnosic acid and tamoxifen.The proliferation ability was measured by MTT assay,the colony formation capacity was measured by colony formation assay,the invasion ability was measured by Transwell chamber assay,the migration ability was measured by wound-healing assay,the expression of Caspase-8,Caspase-3 and PARP protein were measured by Western blotting.Results CA,TAM and CA+TAM suppressed breast cancer cell proliferation with a dose-dependent manner.Compared with CA group and TAM group,the proliferation inhibitory rate of CA+TAM group was significantly increased(P<0.05).In MCF-7 and T47D cell lines,compared to control group,the rate of colony fromation,number of invasive cells and cell migration rate of CA,TAM,CA+TAM groups were significantly decreased(all P<0.05).These indexes mentioned above in CA+TAM group were significantly lower than those in CA group and TAM group(all P<0.05),but there were no significant differences between CA group and TAM group (all P>0.05).In two breast cancer cell strains,the expression of cleaved Caspase-8 protein between control,CA,TAM and CA+TAM groups was not significantly different(P>0.05).Compared to control group,the expression levels of cleaved caspase-3,cleaved PARP protein in CA,TAM and CA+TAM groups were significantly increased(P<0.05),and these protein levels were significantly higher in CA+TAM group than in CA and TAM group(all P<0.05).But there were no significant differences between CA group and TAM group(all P>0.05).Conclusion Carnosic acid combined with tamoxifen can suppress the proliferation,invasion and migration of breast cancer cells,and its mechanism may be related to the activation of Caspase-3/PARP signaling pathway.