他莫昔芬对乳腺癌MCF-7细胞侵袭能力、MMP-9活性和表达的影响及机制  

Effects of tamoxifen on invasion,activity and expression of MMP-9 in breast cancer MCF-7 cells

作　　者：陈妍[1] 王婧[1] 徐旖旎[1] 洪端阳 潘迪[1] 沈祥春[1] 

机构地区：[1]贵州医科大学,贵阳550025

出　　处：《山东医药》2016年第26期6-9,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81302804);贵州省科学技术基金资助项目(黔科合J字20142007号);贵州省高校优秀科技创新人才支持计划项目(黔教合KY字2015492);贵州省中医药管理局中医药;民族医药科学技术研究课题(20141001)

摘　　要：目的观察他莫昔芬(TAM)对乳腺癌MCF-7细胞侵袭能力、基质金属蛋白酶9(MMP-9)活性、MMP-9蛋白表达、MMP-9 mRNA表达的影响,并探讨抑制G蛋白偶联受体30(GPR30)对上述作用的影响。方法取对数生长期乳腺癌MCF-7细胞,用无血清无酚红高糖DMEM培养基培养24 h耗竭内源性雌激素。将MCF-7细胞接种6孔板,待细胞贴壁后分为对照组、TAM组和TAM+G15(GPR30选择性抑制剂)组。对照组用无血清无酚红高糖DMEM培养基继续培养24 h。TAM组用含1μmol/L TAM的无血清无酚红高糖DMEM培养基培养24 h,TAM+G15组先用含1μmol/L的G15的无血清无酚红高糖DMEM培养基培养30 min,然后用含1μmol/L TAM的无血清无酚红高糖DMEM培养基培养24 h。收集各组细胞,用Transwell侵袭实验检测MCF-7细胞侵袭能力,用明胶酶谱法检测MCF-7细胞培养液上清MMP-9活性,用Western blot法检测MCF-7细胞MMP-9蛋白表达,用实时荧光定量PCR法检测MCF-7细胞MMP-9 mRNA表达。结果与对照组相比,TAM组MCF-7细胞侵袭能力升高,MMP-9蛋白和mRNA表达增加,培养液上清MMP-9活性升高(P均<0.05)。与TAM组相比,TAM+G15组上述指标均降低(P均<0.05)。结论 TAM可增强乳腺癌MCF-7细胞的侵袭能力,上调MMP-9的活性及表达水平。用GPR30选择性抑制剂G15抑制GR30通路可以抑制这一作用。Objective To explore the effects of tamoxifen （TAM）on the invasion,activity and expression of matrix met-alloproteinase-9 （MMP-9）in breast cancer MCF-7 cells,meanwhile to discuss the role of inhibiting G-protein-coupled receptor 30 （GPR30）in these effects.Methods MCF-7 cells in the logarithmic phase were pre-cultured for 24 h in phenol red （PR）-free medium without serum to remove endogenous estrogen before the indicated treatments.MCF-7 cells were seeded in the six-well plates and incubated over night to let them adhere on the plate.The control group was continued to cultivate in PR-free me-dium without serum for 24 h.The TAMgroup was treated with 1 μmol/L TAMin PR-free medium without serum for 24 h.And the TAM＋G15 group was pretreated with G15 （1 μmol/L）for 30 min before treatment with TAM（1 μmol/L）for 24 h in PR-free medium without serum.After treatment,cell invasion was detected by Transwell assay.The activity of MMP-9 in culture me-dium was tested by Gelatin zymography assay.The MMP-9 protein expression was analyzed by Western blotting.The mRNA ex-pression of MMP-9 was tested by real-time RT-PCR.Results Compared with control group,the cell invasion was increased, the protein and mRNA expression level of MMP-9 was up-regulated,and the activity of MMP-9 in MCF-7 cells was increased in the TAMgroup （all P 〈0.05）.Furthermore,compared with the TAMgroup,the above indexes of the TAM＋G15 group were all decreased （all P 〈0.05）.Conclusions TAMpromotes the cell invasion ability and up-regulates the activity and expression of MMP-9.Meanwhile,the effects may be suppressed by G15 pretreatment.

关 键 词：他莫昔芬 乳腺癌 基质金属蛋白酶9 G蛋白偶联受体30 

分 类 号：R737.9[医药卫生—肿瘤]