上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

CD147基因沉默抑制三阴性乳腺癌细胞增殖的作用机制

汪成 1, 2，庄志刚 3，单鸣 1 ，徐明 4   

  1. 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
  • 出版日期:2017-03-28 发布日期:2017-03-30
  • 通讯作者: 庄志刚，电子信箱：zhuang_zg@163.com。
  • 作者简介:汪成（1975—），男，副主任医师，博士生；电子信箱：wangcheng1212@hotmail.com。
  • 基金资助:

    上海市卫生与计划生育委员会科研课题（201540259）

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

WANG Cheng1, 2, ZHUANG Zhi-gang3, SHAN Ming1, XU Ming4   

  1. 1. Department of Breast Surgery, The Central Hospital of Huangpu District of Shanghai, Shanghai 200002, China; 2.Tongji University School of Medicine, Shanghai 200092,China; 3. Department of Breast Surgery, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200050, China; 4. Department of Pathology, The Central Hospital of Huangpu District of Shanghai, Shanghai 200002, China
  • Online:2017-03-28 Published:2017-03-30
  • Supported by:

    Project of Shanghai Municipal Health and Family Planning Commission, 201540259

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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 mRNA和蛋白表达。设计针对人CD147基因编码区的siRNA序列，构建重组慢病毒Lv-shRNA-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 mRNA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-shRNA-CD147感染组CD147 mRNA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-shRNA-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。

关键词: 三阴性乳腺癌, CD147, 基因沉默, 增殖, 迁移

Abstract:

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.

Key words: triple negative breast cancer, CD147, gene silence, proliferation, migration