摘要
目的探讨Smad4基因沉默对奥沙利铂(OXA)抑制乳腺癌细胞MDA-MB-231侵袭迁移能力的影响及其作用机制。方法采用MTT法检测不同浓度OXA(0,0.25,0.5,1,2,4μmol/L)对MDA-MB-231细胞的增殖抑制作用;Smad4 siRNA转染MDA-MB-231细胞株,并用Western blot检测转染后Smad4蛋白表达;Transwell小室法和细胞划痕实验检测MDA-MB-231细胞侵袭迁移活性;Western blot和ELISA检测细胞中TGF-β和MMP-9蛋白表达和活性的变化。结果 1μmol/L OXA作用于MDA-MB-231细胞24,48,72 h细胞存活率分别是(77.42±3.98)%,(60.38±3.65)%和(40.15±4.37)%。siRNA转染后MDA-MB-231细胞中Smad4的表达明显被抑制(P<0.05);Smad4 siRNA转染可增强OXA抑制MDA-MB-231细胞的侵袭和迁移活性(P<0.05);Smad4 siRNA转染使OXA下调细胞中TGF-β和MMP-9蛋白表达和活性明显增强(P<0.05)。结论下调Smad4的表达可以增强OXA抑制乳腺癌MDA-MB-231细胞侵袭迁移的活性,其机制可能与影响TGF-β/Smad4通路并下调MMP-9表达有关。
Objective To investigate the effect of small interfering RNA-mediated Smad4 knockdown on oxaliplatin（OXA） inhibiting the invasion and migration of breast cancer cell MDA-MB-231 and to explore its mechanism.Methods The viability of MDA-MB-231 cells exposed to different concentrations（0,0.25,0.5,1,2 and 4 μmol/L） of OXA were detected by MTF assay.Western blot was used to detect Smad4 expression in MDA-MB-231 cells transfected with Smad4 siRNA.Transwell small chamber assay and wound healing assay were used to examine the invasion and migration of MDA-MB-231 cells.TGF-β and MMP-9 expression levels and activities were determined using Western blot and ELISA assay.Results The survival rates of MDA-MB-231 cell were 77.42％,60.38％ and 40.15％ after exposed to OXA （1 μmol/L） for 24,48 and 72 h,respectively.The expression level of Smad4 in MDA-MB-231 cells was significantly inhibited after Smad4 siRNA transfection.Smad4 siRNA transfection enhanced the inhibition of OXA on the MDA-MB-231 invasive and metastasic activities（P ＜ 0.05）.Smad4 siRNA transfection enhanced the down-regulation of expression of TGF-β and MMP-9 in MDA-MB-231 cells by OXA（P ＜ 0.05）.Conclusion Smad4 siRNA transfection can enhance the inhibitory effect of OXA on the invasion and migration of MDA-MB-231 cells,which may be related to the regulation via TGF-β/Smad4 pathway and the down-regulation of MMP-9.
出处
《山西医科大学学报》
CAS
2015年第8期756-761,共6页
Journal of Shanxi Medical University
基金
国家自然科学基金资助项目(81000992
81372899)
安徽省教育厅自然科学研究重点资助项目(KJ2012A202)
安徽省自然科学基金资助项目(1508085MH166)
蚌埠医学院科研基金资助项目(Byky1325)
