circPUM1通过调控miR-524-5p表达对结肠癌细胞恶性生物学行为的影响

目的探讨环状RNA PUM1(circPUM1)对结肠癌细胞增殖、凋亡、迁移、侵袭的影响及其分子机制。方法实时荧光定量聚合酶链反应(qRT-PCR)检测结肠癌组织、癌旁组织中circPUM1、微小RNA-524-5p(miR-524-5p)的表达量。体外培养人结肠癌细胞株SW620,分别将si-NC、si-circPUM1、miR-NC、miR-524-5p mimics、si-circPUM1与anti-miR-NC、si-circPUM1与anti-miR-524-5p转染至SW620细胞。甲基噻唑基四唑(MTT)检测细胞增殖;Transwell小室实验检测细胞迁移、侵袭能力;流式细胞术检测细胞凋亡率;双荧光素酶报告实验验证circPUM1是否能够结合miR-524-5p;蛋白免疫印迹法(Western blotting)检测细胞周期蛋白1(Cyclin D1)、p21、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、B淋巴细胞瘤-2(Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bax)表达量。结果结肠癌组织circPUM1的表达水平显著高于癌旁组织(P<0.05),miR-524-5p的表达水平显著降低(P<0.05);干扰circPUM1表达或miR-524-5p过表达后,细胞活力显著降低(P<0.05),迁移细胞数与侵袭细胞数显著减少(P<0.05),细胞凋亡率显著升高(P<0.05),Cyclin D1、MMP-2、MMP-9、Bcl-2蛋白表达显著降低(P<0.05),p21、Bax蛋白表达显著升高(P<0.05);双荧光素酶报告实验证实circPUM1可靶向结合miR-524-5p的作用位点;抑制miR-524-5p表达可减弱干扰circPUM1表达对SW620细胞增殖、凋亡、迁移、侵袭的作用。结论circPUM1可通过海绵吸附miR-524-5p促进结肠癌细胞增殖、迁移、侵袭,抑制细胞凋亡。

Effect of circPUM1 on the malignant biological behavior of colon cancer cells by regulating the expression of miR-524-5p

Objective To investigate the effect of circPUM1 on the proliferation,apoptosis,migration and invasion of colon cancer cells and its molecular mechanism.Methods qRT-PCR was used to detect the expression of circPUM1 and miR-524-5p in colon cancer tissues and adjacent tissues.Human colon cancer cell line SW620 was cultured in vitro.si-NC,si-circPUM1,miR-NC,miR-524-5p mimics,si-circPUM1 and anti-miR-NC,si-circPUM1 and anti-miR-524-5p were transfected into SW620 cells.MTT was used to detect cell proliferation.The Transwell chamber test was used to detect cell migration and invasion.Flow cytometry was used to detect the apoptotic rate.The double luciferase reporting experiment verified whether circPUM1 can bind to miR-524-5p.Western blotting was used to detect the expression of Cyclin D1,p21,MMP-2,MMP-9,Bcl-2,and Bax.Results The expression of circPUM1 in colon cancer tissues was significantly higher than that in adjacent tissues(P<0.05),and the expression of miR-524-5p was significantly reduced(P<0.05).After interference with circPUM1 expression or miR-524-5p overexpression,cell viability was significantly reduced(P<0.05),the number of migrating cells and invading cells was significantly reduced(P<0.05),but the apoptosis rate was significantly increased(P<0.05),the protein expression of Cyclin D1,MMP-2,MMP-9,and Bcl-2 were significantly reduced(P<0.05),but the protein expression of p21 and Bax were significantly increased(P<0.05).The double luciferase reporting experiment confirmed that circPUM1 could target the action site of miR-524-5p.Inhibition of miR-524-5p expression could attenuate the effects of interfering circPUM1 expression on the proliferation,apoptosis,migration,and invasion of SW620 cells.Conclusion circPUM1 can promote the proliferation,migration and invasion of colon cancer cells,and inhibit cell apoptosis through sponge adsorption of miR-524-5p.