丹参酮ⅡA调控凋亡及信号转导与转录激活因子3信号通路抑制人结肠癌细胞增殖及迁移

Tanshinone ⅡA inhibits migration and invasion of colorectal cancer SW480 cells via regulating apoptosis and signal transducer and activators of transcription 3 signaling pathway

目的:观察丹参酮ⅡA(TanⅡA)对人结肠癌细胞(SW480、HCT116细胞)增殖、凋亡与迁移的影响,探究其分子机制。方法:采用人结肠癌SW480及HCT116细胞,随机分为对照组,丹参酮ⅡA(25、50、100μmol/L)处理组,噻唑蓝比色法(MTT)检测细胞增殖,流式细胞术检测细胞凋亡,划痕试验分析细胞迁移。采用蛋白质印迹法(Western blot)检测SW480细胞中凋亡通路血管内皮生长因子(VEGF)、B细胞淋巴瘤/白血病-2(bcl-2)、bcl-2相关X蛋白(bax)蛋白及信号转导与转录激活因子3(STAT3)信号通路STAT3、c-Myc蛋白表达水平。结果:丹参酮ⅡA呈剂量及时间依赖性的方式抑制SW480及HCT116细胞增殖;与对照组比较,丹参酮ⅡA(50、100μmol/L)处理组细胞凋亡率分别增加了23.1%、35.3%及26.8%、34.6%;丹参酮ⅡA(50、100μmol/L)干预后,两组细胞的迁移抑制率分别增加了26.6%、36.5%及39.8%、42.0%;丹参酮ⅡA干预后SW480细胞中细胞凋亡通路中VEGF、bcl-2蛋白表达下调,bax蛋白表达显著上调;而STAT3信号通路中STAT3、p-STAT3、c-Myc蛋白相对表达量显著降低。结论:丹参酮ⅡA可抑制结肠癌SW480及HCT116细胞增殖,促进凋亡,降低迁移能力,其机制可能与下调凋亡通路中VEGF、bcl-2蛋白并上调bax蛋白表达;下调STAT3信号通路中STAT3、p-STAT3、c-Myc蛋白表达相关。

Objective To investigate the roles of TanshinoneⅡA in the apoptosis,invasion,migration of colorectal cancer SW480 and HCT116 cells and explore the underlying mechanism.Methods SW480 and HCT116 cells were treated with different concentrations(25,50 and 100μmol/L)of tanshinoneⅡA.The inhibition of SW480 cells and HCT116 cells was detected with methyl thiazolyl tetrazolium(MTT)assays.SW480 cells and HCT116 cells apoptosis was evaluated in Hoechst staining.The invasion and migration was examined with Transwell assay and wound healing assay.The protein expression of vascular endothelial growth factor(VEGF),B cell lymphoma/leukemia-2(bcl-2),bcl-2 associated X protein(bax)in apoptosis pathway and signal transducer and activators of transcription 3(STAT3),c-Myc in STAT3 signaling pathway was determined using Western blotting.Results TanshinoneⅡA inhibited the proliferation of SW480 cells and HCT116 cells in a time-and dose-dependent manner.The apoptosis rates was increased by 23.1%/35.3%and 26.8%/34.6%in TanshinoneⅡA treated group(50 and 100μmol/L).TanshinoneⅡA(50 and 100μmol/L)treated also inhibited the migration ability of SW480 cells and HCT116 cells by 26.6%/36.5%and 39.8%/42.0%.After tanshinoneⅡA treatment,the expression of VEGF and bcl-2 protein was significantly decreased and bax was largely increased in apoptosis pathway.Meanwhile,tanshinoneⅡA inhibited the expression of STAT3 and c-Myc in STAT3 signaling pathway.Conclusion TanshinoneⅡA inhibits proliferation,migration and induces apoptosis of colorectal cancer SW480 cells via regulating apoptosis and STAT3 signaling pathway.