摘要
目的:探讨微小RNA-381(miR-381)对前列腺癌细胞增殖、迁移与侵袭的影响,并研究miR-381的靶基因及其功能。方法:采用qRT-PCR检测miR-381在人前列腺癌细胞系中的表达水平。采用基因转染方法过表达或敲减miR-381在前列腺癌细胞中的表达,采用MTT法和克隆形成实验检测细胞增殖能力;流式细胞术检测细胞凋亡;划痕损伤实验、Transwell迁移实验和基质胶侵袭实验检测细胞迁移和侵袭能力。采用生物信息学方法预测miR-381靶点,并采用荧光素酶报告基因分析进行验证。结果:miR-381在前列腺癌细胞中低表达。与空白对照组相比,miR-381 mimics组前列腺癌细胞增殖能力明显降低,克隆数显著减少,凋亡明显增加,迁移和侵袭的细胞数减少;miR-381 inhibitor组结果则相反。荧光素酶报告基因分析证实MAP3K2是miR-381中的靶基因。miR-381 mimics下调而miR-381 inhibitor上调前列腺癌细胞中MAP3K2基因和蛋白表达。结论:miR-381通过靶向MAP3K2抑制前列腺癌细胞增殖、迁移和侵袭,发挥抑癌基因作用。
Objective:To investigate the effects of miR-381 on the proliferation,migration and invasion of prostate cancer cells and to explore the target gene of miR-381.Methods:The expression of miR-381 in human prostate cancer cell lines was determined by qRT-PCR.miR-381 was overexpressed or knocked down in prostate cancer cells by gene transfection.MTT assay and colony formation assay were used to detect the effect of miR-381 on the proliferation of prostate cancer cells.The effect of miR-381 on the apoptosis of prostate cancer cells was detected by flow cytometry.The effects of miR-381 on the migration and invasion abilities of prostate cancer cells were detected by wound healing assay,Transwell migration assay and matrigel invasion assay.The target genes of miR-381 were predicted by bioinformatics and further identified by luciferase reporter gene assay.Results:The expression of miR-381 was lower in prostate cancer cells than normal prostate epithelial cells.Compared with blank control group,the cell viability and the number of cell colonies was significantly reduced while the percentage of apoptotic cells was significantly increased in miR-381 mimics group.The number of migrated and invaded cells was also significantly reduced in miR-381 mimics group.On the contrary,miR-381 inhibitor led to the opposite effects.MAP3K2 was identified as a target gene of miR-381 by luciferase reporter gene assay.miR-381 mimics downregulated while miR-381 inhibitor upregulated the expression of MAP3K2 in prostate cancer cells.Conclusion:miR-381 inhibited the proliferation,migration and invasion of prostate cancer cells by targeting MAP3K2.miR-381 plays tumor suppressive role in prostate cancer,and may be a potential target for the diagnosis and therapy of prostate cancer.
作者
王晖
徐雪莹
张徐
WANG Hui;XU Xue-ying;ZHANG Xu(School of Medicine,Jiangsu University,Zhenjiang Jiangsu 212013,China)
出处
《江苏大学学报（医学版）》
CAS
2019年第3期208-215,共8页
Journal of Jiangsu University：Medicine Edition
基金
江苏省卫计委“六个一工程”项目(LGY2017024)
江苏大学高级人才启动基金(13JDG086)
江苏大学“青年骨干教师培养工程”项目
