目的 探讨细胞外基质金属蛋白酶诱导因子(EMMPRIN)基因对小鼠结肠癌细胞CT26侵袭的影响.方法 构建包含EMMPRIN基因编码框的真核表达载体pCMV-HA2-EMMPRlN,通过脂质体转染CT26细胞,经G418筛选,建立稳定表达EMMPRIN基因的CT26细胞株.对照组为CT26小鼠结肠癌细胞,EMMPRIN组为转染EMMPRIN的小鼠结肠癌细胞.通过体外侵袭实验,分析EMMPRIN CT26细胞侵袭力的改变.通过蛋白质印迹、酶联免疫吸附测定及明胶酶谱实验分析过表达EMMPRIN,CT26细胞基质金属蛋白酶2(MMP-2)合成、分泌、激活的变化.结果 当EMMPRIN基因在CT26细胞稳定过表达后,侵袭实验的结果为EMMPRIN组(98±5)个细胞,对照组(48±6)个细胞(P＜0.01).蛋白质印迹、酶联免疫吸附测定及明胶酶谱实验结果证实,EMMPRIN可明显促进CT26细胞MMP-2的分泌及激活,但对CT26细胞MMP-2的合成无影响.结论 EMMPRIN基因可明显促进小鼠结肠癌细胞CT26的侵袭,并可明显促进CT26细胞MMP-2的分泌及激活.%Objective To investigate whether extracellular matrix metalloproteinase inducer(EMMPRIN)can enhance the metastatic ability of murine colon adenocarcinoma cell(CT26).Methods EMMPRIN was over-expressed in CT26 cells through transfecting pCMV-HA2-EMMPRIN into the CT26 cells.Invasion assay was utilized to analyze the invasion of CT26 cells in vitro after EMMPRIN over-expression.The synthesis of matrix metalloproteinase-2(MMP-2) and the secreted and activated MMP-2 were examined by western blot,Enzyme linked immunosorbent assay(ELISA) and zymography.Results After EMMPRIN over-expression,invasion assay showed that invasive ceils were 98±5 in EMMPRIN group and 48±6 in control group(P ＜ 0.01).The secreted and activated MMP-2 was up-regulated in EMMPRIN group cell culture media,but the synthesis of MMP-2 was no different between EMMPRIN group and control group.Conclusions Over-expression of EMMPRIN can enhance the CT26 cell invasion and up-regulate the secreted and activated MMP-2 in CT26 ceils.The results suggest that EMMPRIN may be involved in cancer metastasis and play an important role in promotion of cancer metastasis.
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