CD147基因沉默抑制三阴性乳腺癌细胞增殖的作用机制
网络出版日期: 2017-03-30
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
Online published: 2017-03-30
Supported by
Project of Shanghai Municipal Health and Family Planning Commission, 201540259
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。
汪成 , 庄志刚 , 单鸣 , 徐明 . CD147基因沉默抑制三阴性乳腺癌细胞增殖的作用机制[J]. 上海交通大学学报(医学版), 2017 , 37(3) : 305 . DOI: 10.3969/j.issn.1674-8115.2017.03.006
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
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