miRNA-195在膀胱癌细胞中功能的研究

摘要

Objective To investigate the influence of microRNA (miRNA)-195 on the proliferation,migration and invasion of human bladder cancer cell 5637.Methods Human bladder cancer cell 5637 was transfected with miRNA-195 by cationic liposomes as miRNA-195 group and transfected with the vector as control group.The expression of miRNA-195 was determined by real-time fluorescence quantitation polymerase chain reaction (PCR).Methyl thiazolyl tetrazolium(MTT)assay,cell counting assay and colony formation assay were used to detect the cell proliferation .Cell migration and invasion were determined by wound healling assay,Transwell assay and Matrigel tumour invasion assay. Results Real-time fluorescence quantitation PCR showed that there was high expression of miRNA-195 after transfection of cell 5637.MTT assay,cell counting assay and colony formation assay demonstrated that miRNA-195 can suppress cell 5637 proliferation in vitro,and there were statistical significance between miRNA-195 and control groups (P<0.05).Wound healing assay,Transwell assay and tumour invasion assay showed that miRNA-195 can suppress cell migration and invasion with statistical significance between the 2 groups(P<0.05 ).Conclusions MiRNA-195 has inhibitory ability on the proliferation,migration and invasion of bladder cancer cell 5637 cell.%目的：研究微小RNA（miRNA）-195对人膀胱癌细胞5637增殖、迁移及侵袭能力的影响。方法利用阳离子脂质体方法将miRNA-195转染至人膀胱癌细胞5637（miRNA-195组），同时以只转染病毒载体的5637细胞作为对照组。采用实时荧光定量聚合酶链反应（PCR）检测转染细胞中miRNA-195的表达量。采用噻唑蓝（MTT）法、细胞计数法和集落形成实验测定miRNA-195对5637细胞体外增殖的影响。采用划痕试验、Transwell实验、Matrigel 肿瘤细胞侵袭实验检测miRNA-195对5637细胞迁移、侵袭能力的影响。结果实时荧光定量PCR检测结果表明转染后的5637细胞高表达miRNA-195。MTT法、细胞计数法及集落形成实验证明miRNA-195可以抑制5637细胞在体外的增殖，miRNA-195组和对照组比较差异有统计学意义（P＜0．05）。划痕实验、Transwell实验以及Matrigel肿瘤细胞侵袭实验证明miRNA-195可以抑制5637细胞的迁移和侵袭能力，miRNA-195组和对照组比较差异有统计学意义（P＜0．05）。结论 miRNA-195能够抑制膀胱癌细胞5637在体外的增殖、迁移以及侵袭能力。