沉默内源性RNF4通过激活p53通路诱导乳腺癌细胞凋亡  

Silencing endogenous RNF4 induces apoptosis of breast cancer cells through activating p53 pathway

作　　者：余思茗 张玥 王婷婷[1] YU Si-ming;ZHANG Yue;WANG Ting-ting(Department of Pharmacy,Peking University Shenzhen Hospital,Shenzhen 518036,China)

机构地区：[1]北京大学深圳医院药学部,518036 [2]深圳北京大学香港科技大学医学中心,518036

出　　处：《中国现代药物应用》2021年第14期245-248,共4页Chinese Journal of Modern Drug Application

基　　金：深圳市科技计划项目(项目编号:JCYJ20190809093205533)。

摘　　要：目的环状泛素连接酶环指蛋白4(RNF4)可增强多种致癌转录因子的稳定性及活性,对肿瘤的生长发展至关重要。在乳腺癌患者中,高水平的RNF4 mRNA与患者的低生存率相关。本研究在离体和在体水平上,探讨沉默乳腺癌细胞中内源性RNF4的表达对于肿瘤生长的抑制作用,并探讨相关分子机制。方法用慢病毒载体系统构建稳定沉默RNF4的乳腺癌细胞系,在离体实验中用CCK-8法检测细胞增殖活力,并用Western-Blot法检测RNF4沉默水平和促凋亡蛋白p53的表达;在体方面,建立皮下荷乳腺癌裸小鼠模型,定期测量裸小鼠皮下瘤体积,绘制肿瘤生长曲线,称量瘤体重量以及检测瘤组织中RNF4和p53的表达。结果与对照组相比,稳定沉默内源性RNF4后,肿癌细胞的增殖活性被显著抑制且促凋亡蛋白p53表达升高;在荷乳腺癌裸小鼠模型中,稳定沉默RNF4组小鼠的肿瘤生长缓慢、肿瘤瘤体体积减小、瘤重减轻且瘤组织中p53表达水平升高。结论沉默乳腺癌细胞中内源性RNF4可上调p53的表达,激活p53通路,从而诱导肿瘤细胞凋亡,有效抑制在体肿瘤的生长。Objective The cyclic ubiquitin ligase,ring finger protein 4(RNF4),can enhance the stability and activity of various oncogenic transcription factors,which is essential for growth and development of tumors.In patients with breast cancer,a high RNF4 mRNA level correlates with a poor survival.Thus,in the present study,to explore the inhibitory effect of silencing endogenous RNF4 on tumor growth in vitro and in vivo,and the molecular mechanism involved.Methods Stably silencing RNF4 silenced breast cancer cell lines were constructed by lentiviral vector system.In vitro,cell proliferation activity was detected by CCK-8 assay,and the silence level of RNF4 and the expression of pro-apoptotic protein p53 were examined by Western-blot assay.In vivo,a tumor model in nude mice bearing subcutaneous breast tumors was established.Subcutaneous tumor volume of nude mice was measured regularly to construct a tumor growth curve,and the tumor weight and the expression of RNF4 and p53 in tumor tissue were measured.Results Silencing endogenous RNF4 significantly inhibited the proliferation of breast cancer cells accompanied by increase of p53.Compared with the control group,stably silencing endogenous RNF4 significantly inhibited the proliferation of tumor cells,and elevated the expression of pro-apoptotic protein p53.In the tumor model in nude mice bearing subcutaneous breast tumors,mice in the stably silencing RNF4 group had slow-growth tumor,reduced tumor volume and weight,and elevated expression of p53.Conclusion Endogenous RNF4 in silencing breast cancer cells can increase expression of p53 and activate p53 pathway,consequently,induce apoptosis of tumor cells and effectively suppress the growth of body tumor.

关 键 词：环指蛋白4 p53 乳腺癌 凋亡 裸小鼠肿瘤模型 

分 类 号：R737.9[医药卫生—肿瘤]