LncR-HOXA-AS3 targeting miR-29a affects apoptosis and autophagy of prostate cancer cells
Objective To investigate the effects of long chain non-coding RNA(lncR)HOXA-AS3 and microRNA-29a(miR-29a)on apoptosis and autophagy of prostate cancer(PCa)cells.Methods Human PCa cells(DU-145,LN-CaP,C4-2B)and human normal prostate cells RWPE-1 were cultured in vitro,and RT-qPCR was used to detect the rela-tive expression of lncR-HOXA-AS3 and miR-29a in the cells.Dual luciferase reporter assay was used to verify the targeted regulatory relationship between lncR-HOXA-AS3 and miR-29a.LNCaP cells in the logarithmic growth phase were divided into the si-NC group,si-HOXA-AS3 group,si-HOXA-AS3+anti-miR-NC group and si-HOXA-AS3+ anti-miR-29a group.Si-NC,si-HOXA-AS3,si-HOXA-AS3+anti-miR-NC,si-HOXA-AS3+ anti-miR-29a were transfected into LNCaP cells of the above groups by liposome transfection.The expression changes of lncR-HOXA-AS3 and miR-29a in each group were detected by RT-qPCR,and the apoptosis of cells in each group was detected by flow cytometry,and the expression levels of apoptosis-related proteins Caspase-3,autophagy related proteins LC3Ⅱ,LC3Ⅰ and Beclin 1 in each group were detected by Western blotting.Results Compared with the normal prostate cells,the expression of lncR-HOXA-AS3 significantly increased(P<0.05),while the expression of miR-29a significantly decreased in human PCa cells(P<0.05).Dual lucifer-ase reporter genes showed that lncR-HOXA-AS3 inhibited luciferase activity by targeting miR-29a(P<0.05).Compared with the si-NC group,the lncR-HOXA-AS3 expression of LNCaP cells decreased,and the miR-29a expression increased(both P<0.05),the apoptosis rate significantly increased(P<0.05),and the protein expression levels of Caspase-3,LC3Ⅱ/LC3Ⅰ and Beclin 1 significantly increased in the si-HOXA-AS3 group(all P<0.05).Compared with the si-HOXA-AS3+anti-miR-NC group,the expression of miR-29a significantly decreased(P<0.05),the apoptosis rate significantly de-creased(P<0.05),and the expression levels of Caspase-3,LC3Ⅱ/LC3Ⅰ,and Beclin 1 proteins significantly decreased in the si-HOXA-AS3+anti-miR-29a group(all P<0.05).Conclusion Inhibiting lncR-HOXA-AS3 can target and regulate the miR-29a gene to promote apoptosis and autophagy in PCa cells.
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