MiR-490-3p Inhibits Prostate Cancer Progression By Targeting HnRNPA1

Objective:Prostate cancer(PCa)is one of the most common types of cancer in men with a rapidly increasing incidence in China in recent years.Micro RNA(miRNAs)is considered to be a key participant in the progression of human cancer,including prostate cancer(PCa).The purpose of this study was to explore the role of miR-490-3p / heterogeneous ribonucleic acid protein A1(hnRNPA1)signaling pathway in PCa and to provide a new target for the treatment of prostate cancer.Methods:1.The expression of miR-490-3p in prostate cancer tissues and cell lines(1)The expression of miR-490-3p in prostate cancer and paracancerous tissues was detected by qRT-PCR.(2)The difference of miR-490-3p expression between prostate cancer cell lines and normal prostate cell were detected by qRT-PCR.2.The effects of miR-490-3p on proliferation,apoptosis,migration and invasion of prostate cancer cells in vivo.(1)The effect of miR-490-3p on the proliferation of prostate cancer cells was detected by CCK-8 kit,EDU kit and cell cloning test respectively.(2)Flow cytometry was used to detect the effect of miR-490-3p on apoptosis of prostate cancer cells.(3)The effect of miR-490-3p on the migration ability of prostate cancer cells was detected by Transwell assay.(4)The effect of miR-490-3p on the invasive ability of prostate cancer cells was detected by Matrix glue assay.3.Molecular mechanism of miR-490-3p in Prostate Cancer progression.(1)The target gene of miR-490-3p was predicted by bioinformatics databases(miRanda,Target Scan).(2)The interaction between miR-490-3p and target gene was verified by qRT-PCR,Western Blotting,double fluorescent gene reporting and other experiments.4.The effects of miR-490-3p on tumorigenesisThe effect of miR-490-3p on tumor formation and target genes was verified by subcutaneous tumorigenesis in nude mice.Results:1.In this study,it was found that the expression of miR-490-3p in prostate tissue was down-regulated compared with that in paracancerous tissues.At the same time,we examined the expression of miR-490-3p in four kinds of prostate cancer cells PC3,DU145,LNCa P and C4-2B.The results showed that the expression of miR-490-3p in prostate cancer cell lines was significantly down-regulated compared with normal prostate cell RWPE-1.2.LNCa P and C4-2B cells were transfected with the miR-490-3p inhibitor or mimics.Investigation demonstrated that an miR-490-3p inhibitor promoted the proliferation,migration,invasion and inhibited the apoptosis of LNCa P cells in vitro.However,miR-490-3p mimic reversed these effects.3.Through Target Scan and miRanda prediction software,we found that hnRNPA1 is a potential target gene of miR-490-3p.Through luciferase reporter gene assay,we confirmed that miR-490-3p can target to bind to hnRNPA1 3'-UTR.Western results showed that miR-490-3p could significantly inhibit the protein expression of hnRNPA1 in prostate cancer cell lines.These results confirm that hnRNPA1 is the target gene of miR-490-3p.4.C4-2B cells transfected with miR-490-3p mimics generated smaller tumors during the same time period,and the mean tumor volumes and weights were significantly lower than the control group.We used qRT-PCR to confirm the expression of miR-490-3p in mice tumour tissues,miR?490?overexpressing C4?2B tumor xenografts owned a higher expression level than the control group.We also used western blot to detect the expression of hnRNPA1 in mice tumours,hnRNPA1 expression was decreased in C4-2B group transfected with miR-490-3p mimics in comparison to controls.These results demonstrated the tumor?suppressive effects of miR-490 in prostate cancer.Conclusion:To sum up,miR-490-3p is lowly expressed in prostate cancer tissues and cell lines,and inhibits the proliferation,migration,invasion and promots apoptosis of prostate cancer cells by targeting hnRNPA1.