西黄丸及其主要成分抑制PI3K/Akt/mTOR信号通路促进PC-3荷瘤小鼠前列腺癌细胞的凋亡

Xihuang Pills and its main components inhibit PI3K/Akt/m TOR signaling pathways and promote the apoptosis of prostate cancer cells in PC-3 tumor-bearing mice

目的:评估西黄丸及其主要成分对裸鼠去势抵抗性人前列腺癌PC-3细胞皮下移植瘤的PI3K/AKT/mTOR信号通路及细胞凋亡的影响。方法:通过西黄丸、麝香、牛黄、多西他赛、麝香+牛黄干预动物模型,计算各组抑瘤率及HE染色观察肿瘤细胞形态,采用qPCR方法检测各组PI3K/Akt/mTOR mRNA水平,Western印迹检测各组PI3K/Akt/mTOR通路和Caspase-3、Caspase-9蛋白的表达情况。结果:西黄丸、麝香、牛黄、麝香-牛黄、多西他赛组连续干预14 d后,各组抑瘤率分别为29.67%、5.52%、7.26%、12.88%、6.26%。肿瘤组织HE染色结果显示药物干预后可见凋亡小体形成,以西黄丸组、麝香+牛黄组明显;西黄丸、麝香+牛黄组PI3K/Akt/mTOR mRNA以及磷酸化蛋白的表达水平均明显下调(P<0.01),并促进Caspase-3、Caspase-9的蛋白表达(P<0.01)。结论:西黄丸及麝香、牛黄具有抑制PC-3细胞皮下移植瘤生长的作用,其机制与抑制异常激活的PI3K/Akt/mTOR信号通路,并促进PC3细胞凋亡有关。

Objective:To evaluate the effects of Xihuang Pills(XHP)and its main components on PI3 K,AKT and mTOR signaling pathways and cell apoptosis of castration-resistant human PCa PC-3 cell subcutaneously transplanted tumors in nude mice.Methods:We assigned 36 PC-3 tumor-bearing model mice to six groups of equal numbers to be treated with XHP,musk,calculus bovis(CB),musk+CB and docetaxel,respectively.After 14 days of intervention,we calculated the tumor-inhibition rate in different groups,observed the morphology of the tumor cells by HE staining,determined the levels of PI3 K,Akt and mTOR mRNA by RT-qPCR,and determined the expressions of PI3 K,Akt and mTOR signaling pathways and caspase-3 and caspase-9 proteins by Western blot.Results:After 14 days of medication,the tumor-inhibition rates in the XHP,musk,CB,musk+CB and docetaxel groups were 29.67%,5.52%,7.26%,12.88%and 6.26%,respectively.HE staining showed the formation of apoptotic bodies in the tumor tissues after intervention,especially in the XHP and musk+CB groups.The mRNA and phosphorylated protein expressions of PI3 K,Akt and mTOR were significantly down-regulated(P<0.01),and so were the expressions of caspase-3 and caspase-9 proteins in the XHP and musk+CB groups in comparison with the control(P<0.01).Conclusion:Xihuang Pills,musk and calculus bovis can inhibit the growth of castration-resistant human PCa PC-3 cell subcutaneously transplanted tumors,which is associated with their effects of suppressing the abnormally activated PI3 K,Akt and mTOR signaling pathways and promoting the apoptosis of PCa PC3 cells.