消岩汤抑制WISP2/CCN5基因敲减的MCF-7细胞增殖的分子机制  

The research on mechanism of Xiaoyan decoction inhibiting the proliferation of MCF-7 with knockdown of WISP2/CCN5 gene

作　　者：吕艳[1] 张畅 李小江[3] 郑旭[1] 韩燕燕[1] 高珊 李翀[1] 耿强[1] LYU Yan;ZHANG Chang;LI Xiao-jiang;ZHENG Xu;HAN Yan-yan;GAO Shan;LI Chong;GENG Qiang(Department of Pathology,First Teach ing Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300381,China)

机构地区：[1]天津中医药大学第一附属医院病理科,天津300381 [2]天津中医药大学研究生院,天津301608 [3]天津中医药大学第一附属医院肿瘤科,天津300381 [4]天津中医药大学第一附属医院实验中心,天津300381

出　　处：《广东医学》2020年第19期1964-1969,共6页Guangdong Medical Journal

基　　金：天津市教委科研计划项目(2017KJ160);天津中医药大学第一附属医院“拓新工程”基金科研课题(201904)。

摘　　要：目的观察消岩汤对WISP2/CCN5基因敲减的MCF-7细胞增殖的抑制作用,以及WISP2/CCN5、Skp2和p27Kip1 mRNA和蛋白水平变化,探索消岩汤抑制WISP2/CCN5基因敲降的MCF-7细胞增殖的分子机制。方法WISP2/CCN5 shRNA慢病毒质粒转染MCF-7细胞,构建WISP2/CCN5基因敲减的MCF-7细胞系,采用高、中、低浓度的消岩汤孵育WISP2/CCN5基因敲减的MCF-7细胞24、36及48 h后,检测细胞增殖抑制率、WISP2/CCN5、Skp2及p27Kip1的mRNA和蛋白水平。结果WISP2/CCN5基因敲减的MCF-7细胞增殖抑制率低于空白组和空载组(P<0.05),消岩汤干预组细胞增殖抑制率高于WISP2/CCN5基因敲减组(P<0.05);高浓度消岩汤孵育WISP2/CCN5基因敲减的MCF-7细胞48 h组细胞增殖抑制率高于相同浓度孵育24 h组(P<0.05)。WISP2/CCN5基因敲减组Skp2 mRNA和蛋白水平均高于空白组和空载组(P<0.05,P<0.05),而p27Kip1 mRNA和蛋白水平则低于空白组和空载组(P<0.05,P<0.05)。消岩汤干预组的Skp2 mRNA和蛋白水平低于WISP2/CCN5基因敲减组(P<0.05,P<0.05),而p27Kip1 mRNA和蛋白水平则高于WISP2/CCN5基因敲减组(P<0.05,P<0.05)。结论消岩汤干预WISP2/CCN5基因敲减的MCF-7细胞,在mRNA和蛋白水平下调Skp2表达,上调p27Kip1表达,抑制MCF-7细胞增殖;2.高浓度消岩汤对细胞增殖活性的抑制较显著,孵育时间较长组对细胞增殖活性的抑制较显著。Objective To investigate the inhibition of Xiaoyan decoction on proliferation of MCF-7 cells with WISP2/CCN5 gene knockdown,and both mRNA and protein levels of WISP2/CCN5,Skp2 and p27Kipl,so the mechanism of Xiaoyan decoction inhibiting proliferation of MCF-7 cells.Methods WISP2/CCN5 shRNA lentivirus plasmid was used to establish the WISP2/CCN5 gene knockdown MCF-7 cells.Xiaoyan decoction with different concentration were used to incubate the MCF-7 cells with W1SP2/CCN5 gene knockdown for 24 h,36 h and 48 h.RT-PCR and Western blot were used to evaluate the effect of Xiaoyan decoction on both mRNA and protein levels of WISP2/CCN5,Skp2 and p27Kipl.Results The inhibition on proliferation of MCF-7 cells with WISP2/CCN5 gene knockdown was significantly higher than that of control group(P<0.05),while the proliferation inhibition rate of MCF-7 cells with WISP2/CCN5 gene knockdown incubated with higher concentration of Xiaoyan decoction for 48 h was significantly higher than that of 24 h with same concentration(P<0.05).Compared to control group,Skp2 mRNA and protein levels of WISP2/CCN5 gene knockdown group was significantly increased(P<0.05),while p27Kipl mRNA and protein levels was significantly reduced(P<0.05).Compared to WISP2/CCN5 gene knockdown group,Skp2 mRNA and protein levels of MCF-7 cells with WISP2/CCN5 gene knockdown incubated by Xiaoyan decoction were significantly reduced(P<0.05),while p27Kipl mRNA and protein levels were significantly increased(P<0.05).Conclusion Xiaoyan decoction inhibits the proliferation of MCF-7 cells with WISP2/CCN5 gene knockdown by up-regulation of Skp2 and down-regulation of p27Kipl at both mRNA and protein levels.Higher concentration and longer incubation of Xiaoyan decoction presents more significant inhibition effects on MCF-7 cells with WISP2/CCN5 gene knockdown.

关 键 词：WISP2/CCN5 SKP2 P27KIPL 基因敲减 消岩汤 

分 类 号：R228[医药卫生—中医基础理论] R737.9[医药卫生—中医学]