lncRNA DCST1-AS1靶向miR-874-3p对直肠癌细胞增殖和凋亡的影响

背景长链非编码RNA(long non-coding RNA,lncRNA)在直肠癌(rectal cancer,RC)中异常表达并可能参与肿瘤发生及发展过程,lncRNA DCST1-AS1在肿瘤中表达上调,但其在RC发生及发展过程中的作用机制尚未阐明,假设lncRNA DCST1-AS1在RC细胞中表达水平升高,并可能促进肿瘤发生及发展.目的研究lncRNA DCST1-AS1对RC细胞增殖、凋亡的影响和潜在的分子机制.方法实时荧光定量聚合酶链式反应检测30例RC组织和癌旁组织中lncRNA DCST1-AS1和miR-874-3p RNA的水平,对RC SW1463细胞进行转染,分为si-NC组、si-DCST1-AS1组、miR-NC组、miR-874-3p组、pcDNA组、pcDNA-DCST1-AS1组、si-DCST1-AS1+antimiR-NC组和si-DCST1-AS1+anti-miR-874-3p组,MTT实验和流式细胞术分别检测各组SW1463细胞增殖光密度(optical density,OD)值和凋亡率,Western blot检测细胞中细胞周期蛋白1(CyclinD1)、p21、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)和Bcl-2相关蛋白(Bcl-2-associated X protein,Bax)表达,双荧光素酶报告系统验证lncRNA DCST1-AS1和miR-874-3p的关系.结果与癌旁组织组相比,在RC组织组中lncRNA DCST1-AS1的含量显著升高(P<0.05),miR-874-3p含量显著降低(P<0.05);与对照si-NC和miR-NC组比较,si-DCST1-AS1组和miR-874-3p组RC SW1463细胞的OD值、CyclinD1和Bcl-2蛋白含量均降低,细胞凋亡率、p21和Bax蛋白含量均升高;lncRNA DCST1-AS1靶向负调控miR-874-3p的表达;与si-DCST1-AS1+anti-miRNC组比较,si-DCST1-AS1+anti-miR-874-3p组SW1463细胞OD值、CyclinD1和Bcl-2蛋白含量升高,细胞凋亡率、p21和Bax含量降低.结论lncRNA DCST1-AS1通过靶向miR-874-3p调控RC SW1463细胞增殖和凋亡,lncRNA DCST1-AS1可能是RC潜在的分子靶点.

LncRNA DCST1-AS1 regulates proliferation and apoptosis of rectal cancer cells by targeting miR-874-3p

BACKGROUND Some long non-coding RNAs(lncRNAs)have been demonstrated to be abnormally expressed in rectal cancer(RC)and may be involved in tumorigenesis and development.The expression of lncRNA DCST1-AS1 is upregulated in tumors,but its mechanism of action in the development and progression of RC has not been elucidated.It was hypothesized that the expression level of DCST1-AS1 is increased in RC cells and it may promote tumorigenesis and development.AIM To investigate the effects of DCST1-AS1 on the proliferation and apoptosis of RC cells and the potential mechanism.METHODS The levels of DCST1-AS1 and miR-874-3p in 30 RC tissues and adjacent tissues were measured by quantitative realtime polymerase chain reaction.RC SW1463 cells were divided into different groups and transfected with si-NC,si-DCST1-AS1,miR-NC,miR-874-3p,pcDNA,pcDNA-DCST1-AS1,si-DCST1-AS1+anti-miR-NC,and si-DCST1-AS1+anti-miR-874-3p,respectively.The proliferation and apoptosis of SW1463 cells in each group were measured by MTT assay and flow cytometry,respectively.Western blot analysis was carried out to measure the expression levels of CyclinD1,p21,B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)proteins in SW1463 cells.The targeting relationship between DCST1-AS1 and miR-874-3p was validated using a dual-luciferase reporter assay system.RESULTS Compared with tumor adjacent tissues,the level of lncRNA DCST1-AS1 in RC tissues was remarkably increased(P<0.05),while the level of miR-874-3p was significantly decreased(P<0.05).Compared with the si-NC and miR-NC groups,cell proliferation and CyclinD1 and Bcl-2 protein levels were reduced in the si-DCST1-AS1 group and miR-874-3p group,while the apoptosis rate and levels of p21 and Bax were increased.LncRNA DCST1-AS1 targeted and negatively regulated the expression of miR-874-3p.Compared with the si-DCST1-AS1+anti-miR-NC group,cell proliferation and CyclinD1 and Bcl-2 protein levels in the si-DCST1-AS1+anti-miR-874-3p group were increased,while cell apoptosis rate and p21 and Bax protein levels were decreased.CONCLUSION LncRNA DCST1-AS1 regulates the proliferation and apoptosis of SW1463 cells by targeting miR-874-3p.DCST1-AS1 may be a potential molecular target for RC.