槐耳清膏对人前列腺癌VCaP细胞增殖的作用  

Effect of Aqueous Extract of Trametes robiniophila on Proliferation of Human Prostate Cancer VCaP Cells

作　　者：杨爱琳[1] 刘亚鑫 黄惠铭 欧阳里山 谢锦欣 刘东晓 王龙燕 屠鹏飞[1] 胡仲冬 YANG Ai-lin;LIU Ya-xin;HUANG Hui-ming;OUYANG Li-shan;XIE Jin-xin;LIU Dong-xiao;WANG Long-yan;TU Peng-fei;HU Zhong-dong(Modern Research Center for Traditional Chinese Medicine,School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中药学院中药现代研究中心,北京100029

出　　处：《中国实验方剂学杂志》2022年第1期79-84,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：北京市科技新星计划项目(Z191100001119083);中华中医药学会青年人才托举工程项目[CACM-2018-(QNRC2-B05)]。

摘　　要：目的:观察槐耳清膏对人前列腺癌VCaP细胞自噬的影响及对核纤层蛋白B;(Lamin B;)表达的调控作用,并探讨其在VCaP细胞增殖中发挥的作用。方法:采用细胞增殖与活性检测(CCK-8)法检测0,2,4,6,8,10 g·L^(-1)槐耳清膏在不同时间下对人前列腺癌VCaP细胞的增殖抑制作用。采用吖啶橙染色法分析槐耳清膏给药对VCaP细胞中自噬溶酶体形成的影响。借助蛋白免疫印迹法(Western blot)检测槐耳清膏给药后细胞中与自噬密切相关的蛋白微管相关蛋白Ⅰ轻链3(LC3),自噬相关蛋白3(Atg3),自噬相关蛋白5(Atg5),自噬相关蛋白7(Atg7)的表达。采用CCK-8法检测槐耳清膏与自噬抑制剂(Bafilomycin A1)联合给药及槐耳清膏单独给药对细胞增殖的作用。通过Western blot检测槐耳清膏对人前列腺癌VCaP细胞中Lamin B;表达的影响,并采用RNA干扰技术探究Lamin B;在槐耳抗人前列腺癌VCaP细胞增殖中的作用。结果:与空白组比较,槐耳清膏能抑制人前列腺癌VCaP细胞的增殖(P<0.01),且呈时间与浓度依赖性。槐耳清膏给药后可促进VCaP细胞中自噬溶酶体的生成。与空白组比较,槐耳清膏能上调VCaP细胞中与自噬进程密切相关的蛋白LC3-Ⅱ,Atg3,Atg5,Atg7表达水平(P<0.05),槐耳清膏能够诱导VCaP细胞自噬的发生。进一步研究发现抑制自噬能够降低人前列腺癌VCaP细胞对槐耳清膏的敏感性(P<0.05)。与空白组比较,槐耳清膏给药能够抑制人前列腺癌VCaP细胞中Lamin B;蛋白表达(P<0.01),下调Lamin B;蛋白表达,在一定程度上也能够减弱VCaP细胞对槐耳清膏的敏感性(P<0.01)。结论:自噬活化和Lamin B;表达下调可能参与了槐耳清膏对人前列腺癌VCaP细胞的增殖抑制作用,能够为槐耳临床应用提供一定的理论基础。Objective:To investigate effect of aqueous extract of Trametes robiniophila(TRM,Huaier on autophagy of human prostate cancer VCaP cells and Lamin B;expression,so as to uncover its role in the proliferation of VCaP cells. Method: The inhibitory effect of 0,2,4,6,8,10 g·L^(-1) TRM aqueous extract on the proliferation of human prostate cancer VCaP cells at different time points were determined by cell counting kit-8(CCK-8)assay. Acridine orange staining was conducted for analyzing the effect of TRM aqueous extract on the formation of autolysosomes in VCaP cells. After medication,the expression of microtubule-associated protein Ⅰ light chain 3(LC3),autophagy-related protein 3(Atg3),autophagy-related protein 5(Atg5),and autophagy-related protein 7(Atg7)in VCaP cells were detected by Western blot. The effect of TRM aqueous extract alone and its combination with autophagy inhibitor bafilomycin A1 on the proliferation of VCaP cells were assayed by CCK-8 assay. RNA interference technology was used to explore the role of Lamin B;in antiproliferation of VCaP cells by TRM. Result: Compared with the blank group,TRM aqueous extract inhibited the proliferation of human prostate cancer VCaP cells in a time-and concentration-dependent manner(P<0.01).Acridine orange staining showed that TRM aqueous extract promoted the formation of autolysosomes in VCaP cells. As revealed by Western blotting,TRM aqueous extract up-regulated the expression levels of LC3-Ⅱ,Atg3,Atg5,and Atg7 in contrast to those in the blank group(P<0.05). All these indicated that TRM aqueous extract induced the autophagy of VCaP cells. In addition,autophagy inhibition impaired the sensitivity of VCaP cells to TRM aqueous extract(P<0.05). The comparison with the blank group showed that TRM aqueous extract inhibited Lamin B;protein expression in VCaP cells(P<0.01),which in turns weakened the sensitivity of VCaP cells to TRM aqueous extract. Conclusion: TRM aqueous extract inhibited the proliferation of human prostate cancer VCaP cells possibly by inducing autogr

关 键 词：槐耳清膏 人前列腺癌VCaP细胞 自噬 核纤层蛋白B 细胞增殖 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]