【摘要】 目的:探讨抑制人结肠癌细胞HCT-116 micro RNA-21表达降低结肠癌细胞侵袭转移能力作用机制。方法:应用表达反义micro RNA-21的质粒mi RZip21转染HCT-116细胞,检测micro RNA-21的变化,Target Scan和Pic Tar等工具筛选与侵袭转移相关的靶基因,实时定量PCR方法检测转染前后靶基因m RNA的变化,Western blot检测靶基因蛋白表达变化,平皿划痕和Transwell实验观察抑制micro RNA-21与否HCT-116细胞转移和侵袭能力。结果:筛选出4种与侵袭转移相关的micro RNA-21靶基因:TIMP-3,RECK,BMPRⅡ和PCDH17;mi RZip21质粒转染HCT-116后,micro-RNA 21表达水平下降约40%,靶基因TIMP-3和RECK的m RNA及靶蛋白表达上升明显,有统计学意义(P<0.05);平皿划痕和Transwell实验结果表明抑制micro RNA-21后,HCT-116细胞转移侵袭能力明显减弱,差异有统计学意义(P<0.05)。结论:抑制micro RNA-21的表达能够减弱HCT-116细胞的侵袭和转移能力。更多还原

【Abstract】 Objective: To explore the effect on invasion and metastasis of HCT-116 colon cancer cell by downregulating the micro RNA-21 expression. Methods: After plasmid mi RZip21, mi RZip TManti-micro RNA-21 expression lentivector, was used to transfect HCT-116,the expression of micro RNA-21 was assessed. The micro RNA-21 target genes associated with invasion and metastasis were predicted by Target Scan and Pic Tar. The m RNA and protein of taget genes were detected by q RT-PCR and Western blot respectively. The ability of migration and invasion of HCT-116 after transfection were detected by the Scratch assay and Transwell assay. Results: TIMP-3, RECK,BMPR Ⅱ and PCDH17 were the gene targets associated with invasion and metastasis; After the cells were transfected with plasmid mi RZip21,the micro RNA-21 level descended to about 60%,the m RNA and protein level of TIMP-3 and RECK increased significantly(P <0.05); Significant reduction of invasion and migration were observed in anti-micro RNA-21-transfected HCT-116(P <0.05).Conclusion: Inhabiting the micro RNA-21 expression can lessen the invasive and metastatic ability of HCT-116 cells.更多还原