摘要
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 observe the effect of Y-box binding protein-1（YB-1） on proliferation,apoptosis and invasion ability of human breast cancer MDA-MB-231 cells.Methods MDA-MB-231 cells were transfected with recombinant plasmid pGSi2 with YB-1 shRNA and negative control plasmid HK in mediation of liposome respectively,then determined for expressions of YB-1,matrix metalloproteinase-2（MMP-2） and MMP-9 at mRNA and protein levels by RT-PCR and Western blot respectively,for proliferation activity,cell cycle and apoptosis by MTT method and flow cytometry,and for invasion ability by Transwell test.Results Both the expression levels of YB-1 mRNA and protein in MDA-MB-231 cells transfected with pGSi2 were significantly lower than those with HK（P 0.01）.After the expression of YB-1 was silenced,the proliferation of MDA-MB-231 cells was inhibited,while the apoptosis was induced,and the invasion ability as well as expression levels of MMP-2 and MMP-9 decreased significantly（each P 0.05）.Conclusion YB-1 shRNA silenced the expression of YB-1 in MDA-MB-231 cells,inhibited the proliferation and invasion ability of cells as well as expressions of MMP-2 and MMP-9,and induced the apoptosis.
出处
《中国生物制品学杂志》
CAS
CSCD
2012年第3期314-317,323,共5页
Chinese Journal of Biologicals
基金
国家自然科学基金(30872417)
