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

作　　者：龙衍 吴泳蓉 郭垠梅 罗新筠 李仙福 黄甜甜 黄振 柳卓 陈铮甲 周青[2] 田雪飞[1,3] LONG Yan;WU Yong-rong;GUO Yin-mei;LUO Xin-jun;LI Xian-fu;HUANG Tian-tian;HUANG Zhen;LIU Zhuo;CHEN Zheng-jia;ZHOU Qing;TIAN Xue-fei(School of Integrated Traditional Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;Department of Andrology,The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410007,China;Key Laboratory of TCM Prescriptions and Syndromes,Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;Scientific Research Office,The Affiliated Hospital of Hunan Acadermy of Traditional Chinese Medicine,Changsha,Hunan 410006,China.)

机构地区：[1]湖南中医药大学中西医结合学院,湖南长沙410208 [2]湖南中医药大学第一附属医院男科,湖南长沙410007 [3]湖南中医药大学中医方证转化重点实验室,湖南长沙410208 [4]湖南省中医药研究院附属医院科研办,湖南长沙410006

出　　处：《中华男科学杂志》2021年第4期340-346,共7页National Journal of Andrology

基　　金：“湖南省高层次卫生人才‘225’工程培养项目”(湘卫函【2019】196号);湖南省教育厅科学研究项目(19A384);国家自然科学基金区域联合创新项目(U20A20408);国家自然科学基金项目(82074450);湖南省性与生殖健康中医临床医学研究中心资助项目(湘科计2020SK4014);省部共建国家重点实验室培育基地开放基金项目(科函[2020]8号04);湖南省教育厅优秀青年项目(20B452);湖南中医药大学校级科研基金(2019XJJJ031)。

摘　　要：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.

关 键 词：西黄丸 前列腺癌 凋亡 PI3K/AKT/MTOR PC-3细胞系 小鼠 

分 类 号：R737.25[医药卫生—肿瘤]