MicroRNA-375表达对大肠癌细胞生物学特性的影响

摘要

Objective:miRNA-375 lentiviral expression vector was build and transfected into CRC cells to inves-tigate the potential effects of miRNA-375 on biological characteristics of CRC cells.Methods:The expression level of miRNA-375 was examined by using quantitative real-time polymerase chain reaction (qRT-PCR).To get the lentiviral plasmids FUA-ZMCS-EF1-EGFP-miR-375 or inhibitor.Subcell line HCT-116 of stable miRNA-375 overexpression and miRNA-375 inhibitor were successfully builded,then the growth in vitro were observed and the growth curves was drawn.The apoptosis rate was detected by the flow cytometer.The effects of miRNA-375 on the migration ability,the invasive potential possibly and intracellular phosphorylation of ERK of HCT-116 cells were detected by MTT assay,scratch test and the Western blot in vitro respectively.Results:The low expression of miRNA- 375 in CRC cells promote cell growth,inhibite apoptosis,promote the intracellular phosphory lation of ERK and in-crease cell migration and invasive ability.Conclusion:Our study indicates that miRNA-375 likes a tumor metastasis suppressor gene in CRC and provides experimental basis for gene diagnosis and therapy.%目的:研究构建miRNA-375慢病毒表达载体并转染结直肠癌细胞,探讨其对结直肠癌细胞生物学特性的影响.方法:采用qRT-PCR法检测结直肠癌细胞株中miRNA-375表达水平.构建FUA-ZMCS-EF1-EGFP-miR-375/inhibitor慢病毒表达载体,建立稳定过表达miRNA-375及其抑制剂的HCT-116亚细胞系,体外观察细胞生长速度并绘制生长曲线,流式细胞仪检测细胞凋亡率,划痕实验检测miRNA-375对HCT-116细胞迁移能力的影响,Western blot法检测miRNA-375对细胞内ERK磷酸化水平的影响.结果:结直肠癌细胞株中miRNA-375低表达不但能促进细胞生长、抑制细胞凋亡,而且能促进细胞内ERK磷酸化水平、增加细胞迁移力和侵袭力.结论:结直肠癌miRNA-375发挥抑癌基因的作用,为miRNA-375在结直肠癌基因诊断及治疗中的应用提供了实验依据.