鳖甲煎丸对肝癌细胞Bcl-2、Bax表达及IL-6/Stat3信号通路的影响

Effects of Bax,Bcl-2 Expression and IL-6/Stat3 Signal Pathway of Hepatocellular Carcinoma Cells Treated with Biejiajian Pill

目的探讨鳖甲煎丸对H22小鼠肝癌细胞的作用机制,并对其相关信号通路进行研究。方法制造H22肝癌移植瘤模型,对荷瘤鼠瘤重、胸腺指数进行检测;采用酶联免疫吸附测定(Elisa)法对荷瘤鼠血清IL-6检测;免疫组织化学法检测H22细胞中Stat5b蛋白表达;应用实时荧光定量PCR(qRT-PCR)法检测鳖甲煎丸对H22细胞的Bcl-2、Bax和Stat3 mRNA表达的影响,蛋白免疫印迹(Western blot)法检测Bax、Bcl-2、Stat3和p-Stat3的蛋白表达。结果与模型对照组相比,鳖甲煎丸降低H22荷瘤鼠瘤重(P<0.01),提高胸腺指数(P<0.05),降低IL-6的含量(P<0.01),具有剂量依赖性;鳖甲煎丸上调Bax mRNA和蛋白表达(P<0.01),下调Bcl-2 mRNA和蛋白的表达(P<0.01);鳖甲煎丸抑制Stat3 mRNA和Stat3、Stat5b蛋白表达(P<0.01),p-Stat3蛋白表达也下调(P<0.05)。结论鳖甲煎丸可能通过下调Bcl-2,上调Bax基因表达,并抑制IL-6/Stat信号通路,发挥其抗肿瘤作用。

Objective The object was to research the anti-tumor effect of Biejiajian Pill on H22 hepatocellular carcinoma cells and the related signal pathway.Methods H22 bearing mice were made.The tumor weigh and index of thymic were measured.IL-6 of mice was quantified via Elisa.The protein of Stat5b was analysed by immunohistochemistry.Bax and Bcl-2 and Stat3 mRNA expression of H22 were measured by qRT-PCR.The protein of Bax,Bcl-2,Stat3 and p-Stat3 were measured by west-ern blot method.Results Compared with the model groups,Biejiajian Pill decreased the tumor weight and IL-6(P<0.01).Biejiajian Pill increased the thymic index(P<0.05)with the dose dependent.Biejiajian Pill up-regulated Bax mRNA and pro-tein(P<0.01).Biejiajian Pill downregulated Bcl-2 mRNA and protein(P<0.01).Biejiajian Pill downregulated Stat3 mR-NA,Stat3 protein,Stat5b(P<0.01)and p-Stat3 protein expression(P<0.05).Conclusion Biejiajian Pill may downregulate Bcl-2 gene expression and up-regulate Bax gene expression to induce cell death.In the meanwhile,the drug inhibited IL-6/Stat signaling pathway and played the anti-tumor role.