瑞戈非尼对肝癌细胞凋亡及琥珀酸脱氢酶D表达的影响

Effects of Regorafenib on Cell Apoptosis and SDHD Expression in Hepatocellular Carcinoma

目的探讨瑞戈非尼对肝癌细胞凋亡及对琥珀酸脱氢酶D(SDHD)表达水平的影响,分析SDHD对肝癌细胞增殖及瑞戈非尼药物作用的影响。方法通过流式细胞术观察瑞戈非尼对肝癌细胞凋亡的影响,通过实时荧光定量聚合酶链反应(RT-qPCR)及蛋白质印迹法(Western blotting)观察瑞戈非尼对SDHD表达的影响。通过CCK-8实验及Edu实验检测SDHD对肝癌细胞增殖的影响,流式细胞术分析SDHD对瑞戈非尼药物作用的影响。通过Western blotting检测SDHD对AKT信号通路的影响,流式细胞术分析AKT信号通路抑制剂MK2206对SDHD促瑞戈非尼作用下细胞凋亡的影响。结果瑞戈非尼促进肝癌细胞凋亡,促进SDHD表达上调(P<0.05);过表达SDHD抑制肝癌细胞增殖(P<0.05),敲减SDHD促进肝癌细胞增殖(P<0.05);MK2206协同SDHD提高肝癌细胞在瑞戈非尼药物作用下的凋亡率(P<0.01)。结论瑞戈非尼可诱发肝癌细胞凋亡,促进SDHD表达上调;SDHD参与调控肝癌细胞增殖及瑞戈非尼药物作用,提示SDHD可能是肝癌治疗的新靶点。

Objective To explore the role of regorafenib on cell apoptosis and SDHD expression in hepatocellular carcinoma(HCC)and analyze the effects of SDHD in HCC proliferation and the therapeutic effect of regorafenib.Methods Flow cytometry was used to analyze the effect of regorafenib on cell apoptosis.RT-qPCR and Western blotting were used to investigate the mRNA level and protein level of SDHD after regorafenib treatment.The effects of SDHD were analyzed in proliferation,the therapeutic effect of regorafenib and the AKT signaling pathway.Flow cytometry was used to analyze the effect of MK2206 coalescing SDHD overexpression on the apoptosis rate under regorafenib treatment.Results Regorafenib induced cell apoptosis,and the expression of SDHD was increased after 24 or 48 hours of regorafenib treatment(P<0.05).SDHD overexpression inhibited cell proliferation(P<0.05)while SDHD knockdown promoted cell proliferation(P<0.05).Consistently,SDHD overexpression in alliance with MK2206 promoted the apoptosis rate under regorafenib treatment(P<0.01).Conclusion Regorafenib treatment induced cell apoptosis and promoted SDHD expression.SDHD was related to cell proliferation and regorafenib's therapeutic effect,which indicated that SDHD appeared to be a potential drug target for HCC.