二氢杨梅素联合顺铂显著诱导PC3细胞中Noxa和Bim过表达  

Combination of dihydromyricetin and cisplatin significantly induces overexpression of Noxa and Bim in PC3 cells

作　　者：王进峰[1] 舒文云 祝宏斌 邝乾华 WANG Jin-feng;SHU Wen-yun;ZHU Hong-bin;KUANG Qian-hua(Department of Urology,Zhejiang Quhua Hospital,Quzhou 324004,China)

机构地区：[1]浙江衢化医院泌尿外科,浙江衢州324004

出　　处：《中国病理生理杂志》2018年第9期1610-1615,共6页Chinese Journal of Pathophysiology

基　　金：衢州市科学技术局市级科技计划项目(No.20172072)

摘　　要：目的:探讨二氢杨梅素对前列腺癌顺铂化疗的辅助作用并研究其机制。方法:MTT法检测前列腺癌细胞系LNCaP和PC3在不同浓度二氢杨梅素和顺铂处理下的细胞活力。Western blot实验检测顺铂和二氢杨梅素联用对PC3细胞FOXO1、Noxa和Bim表达水平、细胞色素C从线粒体中的释放及caspase-9和caspase-3活化水平的影响。免疫共沉淀实验检测PC3细胞中凋亡蛋白酶激活因子1(Apaf-1)和caspase-9的相互作用。流式细胞术检测PC3细胞的凋亡率。结果:二氢杨梅素辅助治疗可明显提高顺铂对前列腺癌的体外抗肿瘤活性(P<0.05)。二氢杨梅素处理能显著促进PC3细胞FOXO1的表达,转染FOXO1小干扰RNA(siRNA)后,二氢杨梅素对顺铂的辅助治疗效果受到明显抑制。二氢杨梅素联合顺铂能显著诱导PC3细胞中Noxa和Bim的过表达,细胞色素C的释放,Apaf-1和caspase-9的相互作用,caspase-9和caspase-3的活化及凋亡的发生。转染FOXO1 siRNA后,二氢杨梅素联合顺铂对PC3细胞的凋亡诱导途径受到显著抑制。结论:二氢杨梅素可能通过FOXO1-Bim/Noxa途径增强顺铂对前列腺癌细胞的体外杀伤活性。AIM:To investigate the adjuvant effect of dihydromyricetin on cisplatin-based chemotherapy in prostate cancer and its mechanisms.METHODS:The viability of LNCaP and PC3 cells treated with different concentrations of dihydromyricetin and cisplatin was measured by MTT assay.The expression of FOXO1,Noxa and Bim,release of cytochrome C from mitochondria,and activation of caspase-9 and caspase-3 in PC3 cells treated with dihydromyricetin and cisplatin were determined by Western blot.Co-immunoprecipitation was performed to detect the interaction of apoptotic protease-activating factor-1(Apaf-1)and caspase-9 in the PC3 cells.The apoptotic rate of PC3 cells was analyzed by flow cytometry.RESULTS:Adjuvant therapy of dihydromyricetin significantly enhanced the anti-tumor effect of cisplatin against prostate cancer in vitro.Dihydromyricetin significantly promoted the expression of FOXO1 in the PC3 cells.However,transfection with FOXO1 small interfering RNA(siRNA)obviously suppressed the adjuvant effect of dihydromyricetin.Combination of cisplatin and dihydromyricetin significantly induced the overexpression of Noxa and Bim,the release of cytochrome C,the interaction of Apaf-1 and caspase-9,the activation of caspase-9 and caspase-3,and the apoptosis in the PC3 cells.On the other hand,transfection with FOXO1 siRNA obviously suppressed the apoptotic pathway of PC3 cells treated with dihydromyricetin and cisplatin.CONCLUSION:Dihydromyricetin enhances the cytotoxicity of cisplatin against prostate cancer through the FOXO1-Bim/Noxa pathway in vitro.

关 键 词：二氢杨梅素 FOXO1-Bim/Noxa信号通路 顺铂 前列腺癌 凋亡蛋白酶激活因子1 

分 类 号：R737.25[医药卫生—肿瘤] R363[医药卫生—临床医学]