LINC00491靶向调控miR-532-3p对前列腺癌细胞增殖、迁移及侵袭的影响

Effect of LINC00491 on proliferation,migration and invasion of prostate cancer cells by target regulating miR-532-3p

目的:探讨LINC00491对前列腺癌细胞增殖、迁移及侵袭的影响及其可能的作用机制。方法:qRT-PCR检测LINC00491、miR-532-3p的表达量;Pearson法分析前列腺癌组织中LINC00491与miR-532-3p表达量的相关性;体外培养人前列腺癌细胞22RV1,采用脂质体转染法将si-NC、si-LINC00491、miR-NC、miR-532-3p mimics、si-LINC00491与anti-miR-NC(共转染)、si-LINC00491与anti-miR-532-3p(共转染)分别转染至22RV1细胞;CCK-8实验检测细胞增殖;流式细胞术检测细胞周期;Transwell检测细胞的迁移及侵袭;双荧光素酶报告实验与qRT-PCR验证LINC00491与miR-532-3p的靶向调控作用;Western blot检测MMP-2、MMP-9蛋白表达量。结果:前列腺癌组织中LINC00491的表达量高于癌旁组织(P<0.05),而miR-532-3p的表达量低于癌旁组织(P<0.05);LINC00491与miR-532-3p呈负相关(r=-0.858 2,P<0.001);分别转染si-LINC00491与miR-532-3p mimics后细胞存活率和MMP-2、MMP-9蛋白水平降低(P<0.05),S期细胞比例降低(P<0.05),迁移及侵袭细胞数减少(P<0.05),G0/G1期细胞比例升高(P<0.05);LINC00491可靶向调节miR-532-3p表达;共转染si-LINC00491与anti-miR-532-3p可恢复si-LINC00491对细胞生物学行为的作用。结论:干扰LINC00491表达可通过上调miR-532-3p表达抑制前列腺癌细胞的增殖、迁移及侵袭。

Objective:To explore the effect of LINC00491 on proliferation,migration and invasion of prostate cancer cells and its possible mechanism.Methods:qRT-PCR was used to detect expression levels of LINC00491 and miR-532-3p;Pearson method was used to analyze the correlation between LINC00491 and miR-532-3p expression in prostate cancer tissues;human prostate cancer cell 22RV1 was cultured in vitro,and si-NC,si-LINC00491,miR-NC,miR-532-3p mimics,si-LINC00491 and anti-miR-NC(co-transfected),si-LINC00491 and anti-miR-532-3p(co-transfection)were transfected into 22RV1 cells by liposome method;CCK-8 experiment was used to detect cell proliferation;flow cytometry was used to detect cell cycle;Transwell test was used to detect cell migration and invasion;dual luciferase reporter experiment and qRT-PCR were used to verify the targeted regulation of LINC00491 and miR-532-3p.Western blot was used to detect protein expressions of MMP-2 and MMP-9.Results:Expression of LINC00491 in prostate cancer tissue was higher than that in adjacent tissues(P<0.05),while expression of miR-532-3p was lower than that in adjacent tissues(P<0.05);LINC00491 was negatively correlated with miR-532-3p(r=−0.8582,P<0.001);after transfection of si-LINC00491 and miR-532-3p mimics,cell survival rate and protein levels of MMP-2 and MMP-9 were decreased(P<0.05),propor‐tion of S-phase cells was decreased(P<0.05),the number of migration and invasion cells was decreased(P<0.05),while the propor‐tion of cells in G 0/G 1 phase was increased(P<0.05);LINC00491 could target regulate expression of miR-532-3p;co-transfection of si-LINC00491 and anti-miR-532-3p could restore the effect of si-LINC00491 on cell biological behavior.Conclusion:Interfering with expression of LINC00491 can inhibit proliferation,migration and invasion of prostate cancer cells by up-regulating the expression of miR-532-3p.