基于AKT信号通路探讨鳖甲煎丸诱导H22肝癌细胞皮下瘤细胞凋亡的作用机制

Mechanism of regulation of subcutaneous transplanted tumor of H22 mouse hepatoma cells by Biejiajian Pills based on AKT signaling pathway

目的:探讨鳖甲煎丸对H22肝癌细胞皮下瘤的作用及相关机制。方法:以BALB/c小鼠为研究对象,建立H22皮下瘤模型,分别用0.9%氯化钠溶液、鳖甲煎丸、环磷酰胺、鳖甲煎丸+环磷酰胺干预小鼠,监测小鼠皮下瘤生长情况。TUNEL染色检测皮下瘤细胞凋亡情况;免疫组化检测cleaved-caspase3的表达;Western Blot法检测cleavedcaspase3、cleaved-caspase9、Bax、Bcl-2、AKT、p-AKT、REK、p-ERK、JNK、p-JNK的蛋白表达。结果:鳖甲煎丸可以显著抑制皮下瘤生长,联合环磷酰胺抑制效果更加明显,并能改善环磷酰胺引起的肝损害;鳖甲煎丸可以诱导皮下瘤细胞凋亡;与模型组比较,联合用药组cleaved-caspase3、cleaved-caspase9、Bax表达上调,Bcl-2、p-AKT表达下调(P<0.01,P<0.05)。结论:鳖甲煎丸可以通过抑制AKT信号通路,诱导H22皮下瘤细胞凋亡,发挥抗肝癌作用,联合环磷酰胺可以增加抗肿瘤作用,并减轻环磷酰胺的毒副作用。

Objective:To study the effect of Biejiajian Pills(BJJ) on subcutaneous tumor of hepatoma cells of H22 mouse and it’s related mechanism.Methods:Taking BALB/c mice as the research object,the H22 subcutaneous transplanted tumor model was established.The mice were divided into four groups,respectively model(normal saline),BJJ,CTX(cyclophosphamide)and BJJ+CTX group.Subcutaneous tumor growth in mice was monitored.TUNEL was used to detecte subcutaneous tumor cell apoptosis.The expression of cleaved-caspase 3 was detected by immunohistochemistry.The protein levels of cleaved-caspase3,cleaved-caspase9,Bax,Bcl-2,AKT,p-AKT,ERK,p-ERK,JNK,p-JNK were measured by Western Blot.Results:The growth of tumors was inhibited by BJJ.The effect was more obvious in the combined group and it could improve the liver damage caused by CTX.The apoptotic cells were increased in BJJ group.Compared with model group,the levels of cleaved-caspase3,cleavedcaspase9,Bax were upregulated in BJJ+CTX group,while Bcl-2,p-AKT were downregulated(P<0.01,P<0.05).Conclusion:BJJ could induce the apoptosis of subcutaneous transplanted tumor cells of H22 mouse hepatoma cells by inhibiting the AKT signaling pathway and play an anti-hepatoma effect.Combined with CTX,BJJ could increase the anti-tumor effect and reduce the toxic and side effects of CTX.