慢病毒介导shRNA抑制IRF-4结合蛋白对乳腺癌细胞生物学特性的影响  

Effects of lentivirus mediated IRF-4 binding protein silencing on biological behavior of breast cancer cells

作　　者：粟薇 杨明珍 张竹君 

机构地区：[1]重庆市北碚区妇幼保健院检验科,400700 [2]第三军医大学第一附属医院医学检验系,重庆400038

出　　处：《免疫学杂志》2016年第3期199-203,共5页Immunological Journal

基　　金：国家自然科学基金(81302316)

摘　　要：目的慢病毒介导sh RNA抑制乳腺癌细胞中免疫相关分子IRF-4结合蛋白(interferon regulatory factor-4 bindingprotein,IBP)的表达,观察对乳腺癌细胞生物学特性的影响。方法针对IBP基因的4个靶点设计干扰序列,干扰慢病毒感染乳腺癌细胞构建稳定感染细胞株;Western blot检测IBP表达水平;相差显微镜观察IBP对乳腺癌细胞形态的影响;MTT实验检测IBP对细胞增殖能力的影响;Transwell实验检测IBP对肿瘤细胞迁移能力的影响;裸鼠动物模型观察IBP对乳腺癌细胞皮下成瘤和肝转移能力的影响。结果选择的干扰靶点均有效,成功构建IBP抑制的乳腺癌细胞株;IBP影响乳腺癌细胞形态,抑制IBP能降低肿瘤细胞增殖和迁移能力,同时降低肿瘤细胞的皮下成瘤和肝转移能力。结论慢病毒介导sh RNA有效抑制了乳腺癌细胞中IBP的表达,降低了肿瘤细胞的增殖和转移能力,IBP有望成为研究肿瘤免疫的新靶点。Our purpose is to silence immune molecular interferon regulatory factor-4 binding protein （IBP） in breast cancer cell via lentivirus assay, and observe its impact on biological behavior of cancer cells. Four target sequences were selected and cloned into lentiviral vector with recombined lentivirus and stable clone was selected pGL V10/U6/RFP/Puro. Breast cancer cells were infected with puromycin, then the expression of IBP was detected by Western blotting and the cell phenotype was observed with phase contrast microscope. MTF assay and transwell chamber assay were performed to investigate the effect of IBP silencing on proliferation and migration of breast cancer cell, respectively. Nude mouse subcutaneous tumor formation and tail veins injection metastatic model were involved to determine the influence of IBP silencing on tumorigenicity and metastasis in vivo. Data showed that all recombinant lentivirus could effectively suppress IBP in breast cancer cell. Phase contrast microscope analysis found that IBP affected the phenotype of breast cancer cells. Suppressing of IBP could significant reduce the proliferation and migration of breast cancer cell in vitro. Nude mice model suggest that IBP silencing dramatically inhibited the tumorigenicity and metastasis of cancer cell in vivo. Taken together, Lentvirus-mediated RNAi technology effectively suppresses IBP in breast cancer cell, and IBP silencing inhibits cell proliferation, migration and metastasis of cancer cells both in vitro and in vivo. Thus, IBP may become a new target for demonstrating tumor immunity and metastasis mechanism.

关 键 词：IRF-4结合蛋白 乳腺癌 转移 慢病毒 

分 类 号：R737.9[医药卫生—肿瘤]