Influences of LncRNA HOXA-AS3 on the proliferation and apoptosis of prostate cancer cells by regulating USP3 via miR-218-5p
Objective To investigate the influences of long non-coding RNA HOXA cluster antisense RNA 3(LncRNA HOXA-AS3)on the proliferation and apoptosis of prostate cancer cells by regulating ubiquitin-specific peptidase 3(USP3)through miR-218-5p.Methods Human prostate epi-thelial cells and human prostate cancer cell lines DU-145,PC-3,22RV1 were cultured in vitro,DU-145 cells were separated into control group,LncRNA HOXA-AS3 overexpression group,LncRNA HOXA-AS3 knockdown group,co-transfection negative control group,and LncRNA HOXA-AS3 knockdown+miR-218-5p inhibitor group.The expression of LncRNA HOXA-AS3,miR-218-5p and USP3was detected by real-time quantitative polymerase chain reaction(PCR)and western blot.The expression ratio of apoptosis-related proteins Bax and Bcl-2(Bax/Bcl-2)was detected by immunofluo-rescence staining.The expression of proliferation(PCNA,Cyclin D1)and apoptosis-related proteins(Bax,Bcl-2)was detected by Western blotting.Detection of targeted regulation of LncRNA HOXA-AS3 on miR-218-5p in prostate cancer cells by double luciferase reporter gene assay.Results Com-pared with human prostate epithelial cells,the protein and mRNA expression of LncRNA HOXA-AS3 and USP3 in DU-145,PC-3 and 22RV1 cells were increased,the expression of miR-218-5p was de-creased(P＜0.05).Compared with the control group,the protein and mRNA expression of LncRNA HOXA-AS3 and USP3,cell viability,colony formation rate,PCNA,and the protein expression of Cyc-lin D1 and Bcl-2 in LncRNA HOXA-AS3 overexpression group were increased,the expression of miR-218-5p,proportion of apoptotic cells,apoptosis rate,Bax/Bcl-2,and the protein expression of Bax were decreased(P＜0.05).The protein and mRNA expression of LncRNA HOXA-AS3 and USP3,cell viability,colony formation rate,PCNA,and the protein expression of Cyclin Dl and Bcl-2 in Ln-cRNA HOXA-AS3 knockdown group were decreased,the expression of miR-218-5p,proportion of ap-optotic cells,apoptosis rate,Bax/Bcl-2,and the protein expression of Bax were increased(P＜0.05).Compared with the LncRNA HOXA-AS3 knockdown group,the protein and mRNA expression of USP3,cell viability,colony formation rate,PCNA,and the protein expression of Cyclin D1 and Bcl-2 in the LncRNA HOXA-AS3 knockdown+miR-218-5p inhibitor group were increased,the ex-pression of miR-218-5p,proportion of apoptotic cells,apoptosis rate,Bax/Bcl-2,and the protein ex-pression of Bax were decreased(P＜0.05).Co-transfection of miR-218-5p mimics and wild-type Ln-cRNA HOXA-AS3 reported a decrease in relative luciferase activity of the plasmid(0.34±0.06 vs.1.01±0.22,P＜0.05).Conclusions Knockdown of LncRNA HOXA-AS3 can reduce the expres-sion of USP3 by up-regulating miR-218-5p,thereby inhibiting the proliferation of prostate cancer cells and promoting their apoptosis.
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