论文部分内容阅读
目的 :研究Semaphorin 4D(Sema 4D)表达沉默对乳腺癌细胞增殖、周期及迁移的影响,初步探讨该蛋白在乳腺癌发生和发展中的作用。方法 :实时荧光定量PCR和蛋白质印迹法检测Sema 4D在不同转移潜能的乳腺癌细胞株MDA-MB-231和MCF-7中的表达水平。采用短发夹RNA(short hairpin RNA,sh RNA)慢病毒感染法,建立Sema 4D稳定低表达的MDA-MB-231细胞系,然后采用CCK-8法、FCM法、划痕愈合实验及Transwell迁移实验检测细胞增殖、周期分布及迁移能力的变化。结果 :Sema 4D在MDA-MB-231细胞中表达水平明显高于MCF-7细胞(P<0.05);sh RNA慢病毒感染可明显下调MDA-MB-231细胞中Sema4D的表达;与空白对照组比较,Sema 4D干扰组的MDA-MB-231细胞增殖明显被抑制(P<0.05),G0/G1期细胞所占比例明显增高(P<0.05),而细胞迁移能力明显降低(P<0.05)。结论:Sema 4D在高转移潜能的乳腺癌MDA-MB-231细胞中高表达,sh RNA干扰Sema 4D表达可抑制该细胞的增殖和迁移,并阻滞细胞周期于G1期。
OBJECTIVE: To study the effect of Semaphorin 4D (Sema 4D) silencing on the proliferation, cycle and migration of breast cancer cells and to explore the role of Semaphorin 4D in the development and progression of breast cancer. Methods: The expression of Sema 4D in breast cancer cell lines MDA-MB-231 and MCF-7 with different metastatic potentials was detected by real-time fluorescence quantitative PCR and Western blotting. The MDA-MB-231 cell line stably expressing low and stable Sema 4D was established by short hairpin RNA (shRNA) transfection. The cells were stained with CCK-8, FCM, scratch healing assay and Transwell migration The changes of cell proliferation, cycle distribution and migration ability were detected by experiments. Results: The expression of Sema 4D in MDA-MB-231 cells was significantly higher than that in MCF-7 cells (P <0.05); sh RNA lentivirus infection could significantly down-regulate Sema4D expression in MDA-MB-231 cells; (P <0.05). The proportion of cells in G0 / G1 phase was significantly increased (P <0.05), while the migration ability of cells was significantly decreased (P <0.05) . Conclusion: Sema 4D is overexpressed in breast cancer MDA-MB-231 cells with high metastatic potential. Shma interference with Sema 4D expression can inhibit the proliferation and migration of the cells and arrest the cell cycle in G1 phase.