CAJ | 학술논문

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.
목적 탐토세포외기질금속단백매유도인자(EMMPRIN)기인대소서결장암세포CT26침습적영향.방법 구건포함EMMPRIN기인편마광적진핵표체재체pCMV-HA2-EMMPRlN,통과지질체전염CT26세포,경G418사선,건립은정표체EMMPRIN기인적CT26세포주.대조조위CT26소서결장암세포,EMMPRIN조위전염EMMPRIN적소서결장암세포.통과체외침습실험,분석EMMPRIN CT26세포침습력적개변.통과단백질인적、매련면역흡부측정급명효매보실험분석과표체EMMPRIN,CT26세포기질금속단백매2(MMP-2)합성、분비、격활적변화.결과 당EMMPRIN기인재CT26세포은정과표체후,침습실험적결과위EMMPRIN조(98±5)개세포,대조조(48±6)개세포(P＜0.01).단백질인적、매련면역흡부측정급명효매보실험결과증실,EMMPRIN가명현촉진CT26세포MMP-2적분비급격활,단대CT26세포MMP-2적합성무영향.결론 EMMPRIN기인가명현촉진소서결장암세포CT26적침습,병가명현촉진CT26세포MMP-2적분비급격활.
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.