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目的:研究小干扰RNA(small interfering RNA,siRNA)抑制轴突导向蛋白分子(Semaphorin4C,Sema4C)基因的表达对人乳腺癌细胞MDA-MB-231体外迁移、侵袭及增殖的影响,并初步探讨其作用机制。方法:根据Sema4C基因设计序列特异性的siRNA(Serna4C-siRNA),在脂质体介导下转染MDA-MB-231细胞,Western blot方法检测基因封闭效应,利用细胞划痕实验、Transwell小室侵袭实验及CFSE流式细胞仪方法检测细胞转染前后迁移、侵袭及增殖能力的变化,Western blot检测磷酸化AKT(p-AKT)在转染前后细胞中表达的变化。结果:转染Sema4C-siRNA 72小时后,乳腺癌MDA-MB-231细胞株(MDA-MB-231/Si)Sema4C蛋白表达明显下降,与未转染细胞MDA-MB-231相比,MDA-MB-231/Si细胞体外迁移能力减弱,侵袭及增殖能力明显下降;p-AKT表达水平在MDA-MB-231/Si细胞中明显降低。结论:Sema4C-SiRNA转染人乳腺癌MDA-MB-231细胞可下调细胞中Sema4c蛋白表达水平,Sema4C-siRNA对MDA-MB-231细胞的体外迁移、侵袭和增殖有抑制作用,这可能与p-AKT的下调有关
OBJECTIVE: To study the effect of small interfering RNA (siRNA) on the migration, invasion and proliferation of human breast cancer cell line MDA-MB-231 in vitro and to explore its possible role in inhibiting the expression of Semaphorin 4C (Sema4C) Mechanism. Methods: According to Sema4C gene sequence-specific siRNA (Serna4C-siRNA), MDA-MB-231 cells were transfected by lipofectamine. Western blotting was used to detect the gene blocking effect. Cell scratch assay, Transwell invasion assay The changes of migration, invasion and proliferation of cells before and after transfection were detected by experiments and CFSE flow cytometry. The expression of phosphorylated AKT (p-AKT) in cells before and after transfection was detected by Western blot. RESULTS: Sema4C protein expression in breast cancer MDA-MB-231 cells (MDA-MB-231 / Si) was significantly decreased after 72 hours of transfection with Sema4C-siRNA. Compared with untransfected cells MDA- MB-231 / Si cells in vitro migration ability weakened, invasion and proliferation decreased significantly; p-AKT expression in MDA-MB-231 / Si cells decreased significantly. Sema4C-siRNA could inhibit the migration, invasion and proliferation of MDA-MB-231 cells in vitro, which may be related to the expression of Sema4c protein in Sertoli cells of MDA-MB-231 cells. -AKT down related