摘要
目的探讨SHP-1对人乳腺癌细胞株MCF-7侵袭能力的影响。方法 MCF-7细胞株分别瞬时转染SHP-1的特异性siRNA干扰序列;然后分别通过RT-PCR和Western-blot检测MCF-7细胞干扰前后SHP-1基因和蛋白表达变化,并通过RT-PCR检测干扰前后MMP-9基因变化;通过Transwell小室实验检测干扰SHP-1对细胞侵袭能力的影响。结果 RT-PCR和Western-blot证实SHP-1在人乳腺癌细胞株MCF-7中有表达以及SHP-1SiRNA的干扰效率;干扰SHP-1后,MMP-9的表达量上调;Transwell小室实验证实,与MCF-7阴性干扰对照及空白对照细胞相比,siRNA组的侵袭能力显著增强(P<0.001)。结论乳腺癌MCF-7中SHP-1的表达水平高低和该细胞侵袭能力高低呈负相关,其侵袭可能和MMP-9的表达呈正相关。
Objective To investigate the effects of SH2 domain-containing cytosolic protein tyrosine phosphatase(SHP-1) on the invasion of MCF-7 breast cancer cells.Methods Cells were transiently transfected with siRNA of the SHP-1 mRNA,then they were analyzed for the presence of SHP-1 using Real-time PCR and western blot analysis.The expression of MMP-9 was detected by Real-time PCR.Cells were analyzed for cell invasion ability using transwell chamber invasion assay.Results SHP-1 gene and protein were found in MCF-7 cells.It was knocked down about 57% by RNA interference.The MMP-9 was increase accompanied with SHP-1 by RNA interference.Transwell chamber invasion assay revealed that SHP-1 RNA interference significantly enhanced the invasion of siRNA transfected cells(P0.001).Conclusion It appears that the expression of SHP-1 had negative effects on the invasion of MCF-7 cells,and the invasion may be positive association with the expression of MMP-9.
出处
《热带医学杂志》
CAS
2011年第3期253-256,F0003,共5页
Journal of Tropical Medicine
基金
广东省自然科学基金(9151051501000016)