CD147基因沉默抑制三阴性乳腺癌细胞增殖的作用机制  

Mechanisms of inhibiting the proliferation of triple negative breast cancer cell lines by silencing the CD147 gene

作　　者：汪成[1,2] 庄志刚[3] 单鸣[1] 徐明[4] WANG Cheng ZHUANG Zhi-gang SHAN Ming XU Ming(Department of Breast Surgery, The Central Hospital of Huangpu District of Shanghai, Shanghai 200002, China Tongji University School of Medicine, Shanghai 200092, China Department of Breast Surgery, Shanghai First Maternity and InJant Hospital, Tongji University School of Medicine, Shanghai 200050, China Department of Pathology, The Central Hospital of Huangpu District of Shanghai, Shanghai 200002, China)

机构地区：[1]上海市黄浦区中心医院乳腺外科,上海200002 [2]同济大学医学院,上海200092 [3]同济大学附属第一妇幼保健院乳腺外科,上海200050 [4]上海市黄浦区中心医院病理科,上海200002

出　　处：《上海交通大学学报（医学版）》2017年第3期305-311,共7页Journal of Shanghai Jiao tong University:Medical Science

基　　金：上海市卫生与计划生育委员会科研课题(201540259)~~

摘　　要：目的·探讨CD147基因沉默对三阴性乳腺癌细胞的增殖抑制作用及机制。方法·体外培养人正常乳腺上皮细胞HMEC及3株三阴性乳腺癌细胞株(MDA-MB-231、HCC70和T4-2),采用real-time PCR和Western blotting分别检测细胞内CD147 m RNA和蛋白表达。设计针对人CD147基因编码区的si RNA序列,构建重组慢病毒Lv-sh RNA-CD147并感染乳腺癌细胞,设立阴性对照(Lv-NC感染组)和不感染细胞对照(细胞对照组)。采用real-time PCR和Western blotting检测CD147基因沉默效果;采用MTT法和Transwell试验分别检测细胞增殖活性和细胞迁移能力。收集病毒感染后72 h的HCC70细胞,采用Western blotting检测细胞内增殖、迁移、凋亡相关蛋白(β-catenin、MMP2、MMP9和Bax)的表达。结果·3株乳腺癌细胞中CD147 m RNA及蛋白的表达均显著高于HMEC细胞(P<0.01)。与Lv-NC感染组和细胞对照组比较,Lv-sh RNA-CD147感染组CD147 m RNA和蛋白表达下调,细胞增殖活性降低,细胞迁移能力减弱,差异均有统计学意义(P<0.01)。病毒感染后72 h,与相应的Lv-NC感染组和细胞对照组比较,Lv-sh RNA-CD147感染组HCC70细胞内β-catenin、MMP2、MMP9表达降低,而Bax表达增高,差异均有统计学意义(P<0.01)。结论·CD147基因沉默可以抑制三阴性乳腺癌细胞的增殖和迁移。Objective · To explore the inhibitive effect of silencing the CD147 gene on the proliferation of triple negative breast cancer cells and relevant mechanisms. Methods · Normal human mammary epithelial cell line HMEC and three triple negative breast cancer cell lines MDA-MB-231, HCC70 and T4-2 were cultured in vitro. mRNA and protein expressions in cells were measured using realtime-PCR and Western blotting, respectively. A siRNA sequence targeting the coding region of human CD147 gene was designed and used to construct a recombinant lentivirus Lv-shRNA-CD147, which was used to infect the three breast cancer cell lines. The negative control group （Lv-NC infected group） and the noninfected cell control group （cell control group） were simultaneously used. The effects of silencing the CD147 gene were measured with real-time PCR and Western blotting. The proliferation and migration of cells were measured with MTT and Transwell assay, respectively. HCC70 cells were collected 72 h after viral infection and proteins related to proliferation, migration and apoptosis of cells （β-catenin, MMP2, MMP9, and Bax） were measured with Western blotting. Results · mRNA and protein expressions of CD147 were significantly higher in three breast cancer cell lines than in HMEC （P〈0.01）. The Lv-shRNA-CD147 infected group had lower mRNA and protein expressions of CD147, cell proliferation, and cell migration as compared with the Lv-NC infected group and the cell control group, the differences were statistically significant （P〈0.01）. The Lv-shRNA-CD147 infected group had lower expressions of β-catenin, MMP2, and MMP9, and higher Bax expression in HCC70 cells 72h after viral infection as compared with the Lv-NC infected group and the cell control group, the differences were statistically significant （P〈0.01）. Conclusion · Silencing the CD147 gene can inhibit the proliferation and migration of triple negative breast cancer cells.

关 键 词：三阴性乳腺癌 CD147 基因沉默 增殖 迁移 

分 类 号：R737.9[医药卫生—肿瘤]