雌二醇通过TGFβ1促进前列腺间质细胞MMP-2的表达  

Up-regulation of MMP-2 Expression by Estradiol via TGFβ1 in Prostate Stromal Cells

作　　者：苗琳[1] 周颖[1] 石建党[1] 杜小玲[1] 焦鸿丽[1] 张琚[1] 

机构地区：[1]南开大学生命科学学院生物化学与分子生物学系,生物活性材料教育部重点实验室,天津300071

出　　处：《中国生物化学与分子生物学报》2009年第8期712-718,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.30672101,30872592);高等学校博士学科点专项科研基金(No.20070055023);国家重点基础研究发展计划(973基金,No.2009CB918900);天津市科委攻关培育基金(No.07jczdjc08300)资助~~

摘　　要：基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)在前列腺间质细胞中表达,转化生长因子β1(transforming growth factorβ1,TGFβ1)可以通过上调基质金属蛋白酶的表达促进肿瘤细胞迁移.我们近期发现,雌二醇可以诱导原代前列腺间质细胞TGFβ1表达.由此我们提出,雌二醇通过上调TGFβ1促进前列腺间质细胞MMP-2表达的假说.用实时定量RT-PCR和酶谱电泳技术分别检测添加雌二醇、TGFβ1、雌二醇和TGFβ1中和抗体、TGFβ1和放线菌素D或TGFβ1和放线菌酮,对前列腺间质细胞系WPMY-1中MMP-2表达的调节作用及其分子机制;荧光素酶活性实验检测TGFβ1对MMP-2启动子活性的影响.结果显示,雌二醇和TGFβ1均以剂量依赖方式促进WPMY-1中MMP-2蛋白水平表达,而对其mRNA表达没有调节作用.雌二醇可以在转录和翻译水平上促进TGFβ1表达;TGFβ1不能促进MMP-2启动子的活性;雌二醇对MMP-2蛋白表达的促进作用可以被TGFβ1中和抗体阻断.放线菌酮而非放线菌素D可以抑制TGFβ1对MMP-2表达的上调作用.表明雌二醇可以在TGFβ1的介导下促进WPMY-1中MMP-2的表达;TGFβ1对MMP-2表达的促进作用是一种转录后的调节机制.结果提示,雌二醇上调间质细胞MMP-2的表达可能是雌激素参与前列腺癌转移和进展的机制之一.Matrix metalloproteinase-2 （MMP-2） were mainly expressed in prostate stromal cells. Transforming growth factor β1 （TGFβ1） may promote tumor cell migration through increasing the expressions of matrix metalloproteinases （MMPs）. Estradiol induced the expressions of TGFβ1 in prostate primary stromal cells. From this, it was proposed that estradiol increased the expressions of MMP-2 via TGFβ1 in prostate stromal cells. Rea/-time RT-PCR and gelatin zymography were performed to detect the effects and mechanisms of MMP-2 expressions in WPMY-1, a prostatic stromal cell line after stimulated with estradiol, TGFβ1, estradiol and TGFβ1 neutralizing antibody, TGFβ1 and cycloheximide （CHX）, or TGFβ1 and aetinomycin D （ActD）.The luciferase activity assay were performed to examine the effects of TGFβ1 on MMP-2 promoter activities. The results showed that both estradiol and TGFβ1 stimulated the MMP-2 protein expressions in a dosedependent manner, but had no effects on MMP-2 mRNA expressions in WPMY-1. Estradiol also stimulated TGFβ1 mRNA and protein expressions. TGFβ1 had no effects on MMP-2 promoter activities. The up-regulation of MMP-2 protein expression by estradiol was blocked by TGFβ1 neutralizing antibody. CHX, instead of ActD, significantly inhibited the effects of TGFβ1 on the expressions of MMP-2. Our data suggested that estradiol induced the expressions of MMP-2 in WPMY-1 via TGFβ1. The up-regulation of MMP-2 by TGFβ1 is through a post-transcriptional mechanism, by which estrogen may stimulate the migrations of prostate cancer cells and promote prostate cancer progressions.

关 键 词：雌激素 基质金属蛋白酶-2 转化生长因子Β1 前列腺癌转移 

分 类 号：Q5[生物学—生物化学] Q71