CD147基因沉默抑制三阴性乳腺癌细胞增殖的作用机制

发布时间：2018-04-27 09:53

本文选题：三阴性乳腺癌 + CD　；参考：《上海交通大学学报(医学版)》2017年03期

【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(P0.01)。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(P0.01)。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(P0.01)。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。
[Abstract]:Objective to investigate the inhibitory effect and mechanism of CD147 gene silencing on the proliferation of three negative breast cancer cells. Methods Human normal breast epithelial cells (HMEC) and three three-negative breast cancer cell lines (MDA-MB-231) HCC70 and T4-2 were cultured in vitro. The expression of CD147 m RNA and protein were detected by real-time PCR and Western blotting, respectively. Si RNA sequence targeting the coding region of human CD147 gene was designed. The recombinant lentivirus Lv-sh RNA-CD147 was constructed and infected with breast cancer cells. The negative control group (Lv-NC infection group) and the non-infected cell control group (cell control group) were established. The silencing effect of CD147 gene was detected by real-time PCR and Western blotting, and the cell proliferation activity and cell migration ability were detected by MTT assay and Transwell test, respectively. HCC70 cells were collected at 72 h after infection. The proliferation, migration and expression of apoptosis-related proteins (尾 -cateninin MMP2, MMP9 and Bax) were detected by Western blotting. Results the expression of CD147 m RNA and protein in three breast cancer cells was significantly higher than that in HMEC cells. Compared with Lv-NC infection group and cell control group, the expression of CD147 m RNA and protein in Lv-sh RNA-CD147 infection group was down-regulated, cell proliferation activity was decreased, cell migration ability was weakened, and the difference was statistically significant (P 0.01). 72 h after infection, the expression of 尾 -cateninine MMP2mMP9 in HCC70 cells was decreased and the expression of Bax was increased in Lv-sh RNA-CD147 infected HCC70 cells, compared with the corresponding Lv-NC infection group and the cell control group. The difference was statistically significant (P 0.01). Conclusion CD147 gene silencing can inhibit the proliferation and migration of three negative breast cancer cells.
【作者单位】：上海市黄浦区中心医院乳腺外科;同济大学医学院;同济大学附属第一妇幼保健院乳腺外科;上海市黄浦区中心医院病理科;
【基金】：上海市卫生与计划生育委员会科研课题(201540259)~~
【分类号】：R737.9

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