摘要
目的 探讨抑癌基因乳腺癌转移抑制基因1（BRMS1）影响胶质瘤细胞侵袭的机制.方法 将真核表达质粒pEGFP-BRMS1和对照质粒pEGFP-N2分别转入人胶质瘤U251和U87细胞中,Western blot技术检测BRMS1蛋白表达,人工基底膜侵袭实验检测细胞侵袭能力,明胶酶谱实验检测胶质瘤细胞分泌基质金属蛋白酶-2（MMP-2）的活性,Western blot检测金属蛋白酶组织抑制因子-2（TIMP-2）及MMP-2蛋白表达.结果 与对照组比较,转染pEGFP-BRMS1质粒24 h后U251和U87细胞侵袭力分别下降了43％和52％,差异均有统计学意义（P＜0.01）；转染BRMS1可以明显抑制2种胶质瘤细胞分泌MMP-2活性；pEGFP-BRMS1可以通过上调TIMP-2的表达从而抑制MMP-2蛋白表达.结论 BRMS1可以通过增加TIMP-2表达进而抑制MMP-2的表达,打破TIMP-2/MMP-2平衡,最终抑制胶质瘤细胞侵袭能力.
Objective To investigate the effect and mechanism of breast cancer metastasis suppressor-1 （BRMS1） on the human glioma cells invasion.Methods Transfection of the pEGFP-BRMS1 plasmids into the U251 and U87 glioma cells were carried out using Lipofectamine 2000.We studied the role of BRMS1 in glioma cell invasion by matrigel cell invasion assay.We performed gelatin zymography to measure the matrix metalloproteinase （MMP）-2 activities in glioma cells and Western blotting to examine the tissue inhibitorof metalloproteinase-2 （TIMP-2） and MMP-2 proteins expression after BRMS1 restoration.Results Compared to the control plasmids,pEGFP-BRMS1 plasmids could increase the expression of BRMS1 in both glioma cells significantly.In cell invasion assay,BRMS1 inhibits cell invasion ability of U251 and U87 cells in matrigel-coated Boyden chamber by 43% and 52%,respectively.BRMS1 also suppress the MMP-2 activity.Increasing of BRMS1 expression upregulated TIMP-2 protein expression and inhibited MMP-2 protein expression.Conclusion Our data indicate that BRMS1 may be reduced glioma cells invasion abilities by the imbalance between TIMP-2 and MMP-2.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第11期2327-2329,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（81201636）
江苏省自然科学基金资助项目（BK2012146）
