论文部分内容阅读
目的观察Y-盒结合蛋白-1(Y-box binding protein-1,YB-1)基因对人乳腺癌细胞株MDA-MB-231增殖、凋亡及侵袭能力的影响。方法通过脂质体介导的方法将YB-1 shRNA重组质粒pGSi2和阴性对照质粒HK转染MDA-MB-231细胞,采用RT-PCR和Western blot法检测细胞中YB-1、基质金属蛋白酶-2(Matrix metalloproteinase-2,MMP-2)、MMP-9在mRNA和蛋白水平的表达;MTT法及流式细胞术检测细胞增殖活力、细胞周期及凋亡的变化;Transwell小室法检测细胞侵袭能力的变化。结果 pGSi2组MDA-MB-231细胞较HK组细胞YB-1 mRNA及蛋白的表达水平均明显降低(P<0.01);沉默YB-1表达可以抑制MDA-MB-231细胞增殖,诱导其凋亡,且细胞侵袭能力及MMP-2和MMP-9的表达也较HK组明显下降(P均<0.05)。结论YB-1 shRNA可以沉默MDA-MB-231细胞中YB-1的表达,抑制细胞增殖,诱导其凋亡,并抑制细胞侵袭能力及MMP-2和MMP-9的表达。
Objective To investigate the effects of Y-box binding protein-1 (Y-box binding protein-1) on the proliferation, apoptosis and invasion of human breast cancer cell line MDA-MB-231. Methods The YB-1 shRNA recombinant plasmid pGSi2 and the negative control plasmid HK were transfected into MDA-MB-231 cells by liposome-mediated method. The expression of YB-1 and MMP-2 were detected by RT-PCR and Western blot. 2 (MMP-2), MMP-9 at mRNA and protein levels; MTT assay and flow cytometry were used to detect cell proliferation activity, cell cycle and apoptosis; Transwell chamber assay of cell invasion The change. Results The expression of YB-1 mRNA and protein in MDA-MB-231 cells in pGSi2 group was significantly lower than that in HK group (P <0.01). Silencing YB-1 expression could inhibit the proliferation of MDA-MB-231 cells and induce apoptosis , And the cell invasion ability and the expression of MMP-2 and MMP-9 were also significantly lower than those in HK group (all P <0.05). Conclusion YB-1 shRNA can silence the expression of YB-1 in MDA-MB-231 cells, inhibit cell proliferation, induce apoptosis, and inhibit cell invasiveness and MMP-2 and MMP-9 expression.