siRNA沉默Sema 4D对乳腺癌MDA-MB-231细胞增殖及迁移的影响

Effects of siRNA-mediated Semaphorin 4D silencing on proliferation and migration of human breast cancer MDA-MB-231 cells

目的:观察siRNA沉默信号素4D(semaphorin 4D,Sema 4D)对乳腺癌MDA-MB-231细胞增殖、迁移的影响。方法:构建靶向Sema 4D基因的siRNA,脂质体法转染MDA-MB-231细胞系,通过实时荧光定量-PCR、Western blotting检测干扰效率,筛选有效序列。siRNA有效干扰MDA-MB-231细胞Sema 4D表达后,CCK-8法及Transwell迁移实验检测细胞增殖及迁移能力的变化。结果:Sema 4D siRNA转染MDA-MB-231细胞后Sema 4D的(mRNA及蛋白)表达水平明显下降(P<0.05),其中以siRNA-C最明显(P<0.05);与空白、阴性对照组相比,siRNA-C沉默MDA-MB-231细胞Sema 4D表达可明显抑制MDA-MB-231细胞增殖(均P<0.05),同时明显减弱细胞的迁移能力[穿膜细胞数:(105.60±12.07)vs(196.20±9.04)、(186.40±6.69)个,均P<0.05]。结论:siRNA沉默Sema 4D可抑制MDA-MB-231细胞增殖,并抑制细胞迁移,可作为乳腺癌基因治疗的潜在位点。

Objective: To investigate the effects of siRNA-mediated Semaphorin 4D( Sema 4D) gene silence on the proliferation and migration of breast cancer MDA-MB-231 cells. Methods: Small inference RNA targeting different regions of Sema 4D gene were transfected into MDA-MB-231 cells by lipofectamine 2000. The most effective siRNA in knockdown Sema 4D was screened by using quantitative PCR and Western blotting. The changes of cell proliferation and migration ability after Sema 4D knockdown were examined by CCK-8 and transwell assays. Results: MDA-MB-231 cells transfected with Sema 4D siRNA had significantly decreased expression of Sema 4D,especially in these transfected with siRNA-C( P < 0. 05). Compared with the blank and scrambled siRNA transfection groups,siRNA-C transfection significantly inhibited the proliferation of MDA-MB-231 cells( P < 0. 05) and their migration ability( numbers of migration cell:[196. 20 ± 9. 04]and [186. 40 ± 6. 69]vs [105. 60 ± 12. 07],P < 0. 05). Conclusion: Down-regulation of Sema 4D by siRNA significantly inhibits the proliferation and migration of MDA-MB-231 cells,suggesting Sema 4D is a novel target for breast cancer therapy.