慢病毒介导的细胞分裂周期相关蛋白2基因沉默对乳腺癌细胞增殖和凋亡的影响

目的 探讨应用慢病毒介导小干扰核糖核酸(siRNA)沉默细胞分裂周期相关蛋白2(CDCA2)基因对乳腺癌细胞增殖和凋亡的影响.方法 采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)与蛋白质印迹法(Western blotting)法分别检测人正常乳腺上皮细胞株MCF-10A与乳腺癌细胞MCF-7、MDA-MB-23...

Effect of CDCA2 gene inhibition by lentivirus-mediated small interfering RNA on proliferation and apoptosis of breast cancer cells

Objective To investigate the effects of lentiviral-mediated small interfering ribonucleic acid (siRNA) silencing of cell division cycle-associated protein 2 (CDCA2) gene on proliferation and apoptosis of breast cancer cells.Methods The expressions of CDCA2 mRNA and protein in human normal mammary epithelial cell line MCF-10A and breast cancer cells MCF-7, MDA-MB-231 and BT549 were detected by qRT-PCR and Western blotting. The lentiviral expression vector was constructed and transfected into breast cancer MCF-7 cells. The experiment was assigned into Control group, Lv-NC group and Lv-siRNA-CDCA2 group. The effects of silencing CDCA2 expression on cell proliferation were detected by Methyl Thiazolyl Tetrazolium (MTT) assay and colony formation assay.The effect of silencing CDCA2 expression on apoptosis was detected by flow cytometry. The expression of Cyclin D1, CDK4, P21, Bcl-2 and Bax protein was detected by Western blotting. The effect of Lv-siRNA-CDCA2 on the volume and weight of transplanted tumor was observed by nude mice xenograft experiments.Results The expression levels of CDCA2 mRNA [(2.18±0.13), (1.56±0.09), (1.83±0.10) vs. (1.02±0.06)] and protein [(1.13±0.15), (0.56±0.08), (0.79±0.11) vs. (0.23±0.04)] in breast cancer cells MCF-7, MDA-MB-231 and BT549 were significantly higher than those in MCF-10A (P<0.05), and the highest level of CDCA2 expression was observed in breast cancer MCF-7 cells. Compared with Lv-NC group, the proliferation activity and colony formation rate of MCF-7 cells in Lv-siRNA-CDCA2 group were significantly decreased (P<0.05), while the apoptosis rate was significantly increased (P<0.05), P21 and Bax expression were promoted (P<0.05), and Cyclin D1, CDK4 and Bcl-2 expression were inhibited (P<0.05). Compared with the Lv-NC group, the volume and weight of the transplanted tumors in the Lv-siRNA-CDCA2 group were significantly lower (P<0.05).Conclusion Lentiviral-mediated CDCA2 gene silencing inhibits breast cancer cells proliferation, induces apoptosis, and inhibits tumor growth.