miR-1229-5p对人结直肠癌细胞SW480增殖、迁移及侵袭的作用研究

Effect of miR-1229-5p on proliferation,migration and invasion of human colorectal cancer cells SW480

目的:探讨miR-1229-5p对SW480细胞生物学功能的影响和作用机制。方法:采用瞬时转染在SW480细胞中过表达miR-1229-5p,进行细胞克隆、EdU实验、MTT、细胞划痕、Transwell小室迁移及侵袭实验,检测miR-1229-5p mimics组与对照组结直肠癌细胞SW480生物学功能变化。转染过表达miR-1229-5p慢病毒后进行裸鼠皮下成瘤实验,观察miR-1229-5p在体内对细胞增殖能力的影响。通过预测软件TargetScan筛选出X连锁凋亡抑制蛋白相关因子1(XAF1)作为miR-1229-5p的潜在下游靶基因,并检测miR-1229-5p对其蛋白表达的影响。在SW480细胞中转染XAF1过表达质粒,共转染miR-1229-5p后进行回复实验验证miR-1229-5p通过靶向XAF1促进SW480细胞增殖及侵袭。结果:miR-1229-5p mimics组较miR-NC组细胞的增殖、迁移及侵袭能力均提高(P<0.01);miR-1229-5p组瘤体体积高于miR-NC组(P<0.01);miR-1229-5p组对靶基因XAF1表达量低于miR-NC组(P<0.01),转染XAF1质粒后可逆转miR-1229-5p mimics对细胞增殖及侵袭的促进作用(P<0.01)。结论:miR-1229-5p可通过靶向XAF1促进结直肠癌细胞SW480的增殖和侵袭能力。

Objective:To investigate the effect of miR-1229-5p on the biological function of SW480 cells and the mechanism of action.Methods:Transient transfection was used to overexpress miR-1229-5p in SW480 cells,and cell cloning,EdU assay,MTT,cell scratching transwell migration and invasion assays were performed to detect the effects of miR-1229-5p mimics group and control group on the biological functions of colorectal cancer cells SW480.Subcutaneous tumorigenesis assay was performed in nude mice after transfection with miR-1229-5p lentivirus to observe the effect of miR-1229-5p on cell proliferation ability in vivo.In addition,X-linked inhibitor of apoptosis associated factor 1(XAF1)was screened by predictive software as a potential downstream target gene of miR-1229-5p and the effect of miR-1229-5p on its protein expression was examined.The XAF1 overexpression plasmid was transfected in SW480 cells,and miR-1229-5p was cotransfected with miR-1229-5p to verify that miR-1229-5p promoted the proliferation and invasion of SW480 cells by targeting XAF1.Results:The proliferation,migration and invasion ability of miR-1229-5p mimics group were increased compared with miR-NC group(P<0.01).The tumor volume of miR-1229-5p group was higher than that of miR-NC group(P<0.01).The expression level of target gene XAF1 in miR-1229-5p group was lower than that in miR-NC group(P<0.01),and the promotion effect of miR-1229-5p mimics on cell proliferation and invasion could be reversed after transfection of XAF1 plasmid(P<0.01).Conclusions:miR-1229-5p can promote the proliferation and invasion of colorectal cancer cells SW480 by targeting XAF1.