摘要
Abstract
Objective To explore the effects of miRNA-17-5p (miR-17-5p) on human colon carcinoma cells proliferation, invasion and migration, and to identify the target gene and ability of miR-17-5p. Methods The miR-17-5p expressions in colon carcinoma and adjacent normal tissues were detected using real-time fluorescent quantified PCR (polymerase chain reaction). SW620 and SW480 cell lines were respectively transfected with miR-17-5p mimics, miR-17-5p antisense oligonucleotide (ASO) and miRNA controls. Cell proliferation ability was detected using methyl thiazolyl tetrazolium (MTT) and clonality methods, and the cell invasion and migration abilities in vitro were detected using Tran-swell chamber kit. The bioinformatics was used to predict the target gene of miR-17-5p, and then enhanced green fluores-cent protein (EGFP) reporter, real-time fluorescent quantified PCR and Western blot methods were used to validate the target gene. Finally, overexpression plasmid pcDNA3-PTEN in target gene were transfected to cells to detect the prolifera-tion, invasion and migration abilities in vitro. Results MiR-17-5p showed an increased expression in the colon carcino-ma tissues compared with that in the adjacent tissues (P < 0. 01). The SW620 and SW480 cytoactive, clonality, invasion and migration abilities after miR-17-5p transfection were enhanced compared with those in control group (P < 0. 01), while the SW620 and SW480 cytoactive, clonality, invasion and migration abilities were decreased after blocking miR-17-5p expression by ASO (P < 0. 01). Overexpression miR-17-5p could inhibit the intensity of EGFP reporter supportor on miR-17-5p combining site (P < 0. 01), and the inhibition became weakened after combining site mutation. The overex-pression of miR-17-5p inhibited PTEN expression on both mRNA and protein levels (P < 0. 01), while blocking miR-17-5p expression by ASO could increase PTEN expression (P < 0. 01). Overexpression of PTEN reduced the SW480 and SW620 cytoactive, clonality, invasion and migration abilities (P < 0. 01). Conclusion PTEN is a direct target gene of miR-17-5p, and miR-17-5p may promote the cell proliferation, invasion and migration abilities by inhibiting PTEN ex-pression.关键词
微RNAs/结肠肿瘤/细胞增殖/PTEN 磷酸水解酶Key words
MicroRNAs/Colonic neoplasms/Cell proliferation/PTEN phosphohydrolase分类
医药卫生
