SLC16A1-AS1负调控miR-450b-5p对结直肠癌细胞Caco-2细胞周期、迁移、侵袭和凋亡的影响

Effects of SLC16A1-AS1 on cell cycle,migration,invasion and apoptosis of colorectal cancer cell Caco-2 by negatively regulating miR-450b-5p

目的探讨SLC16A1-AS1对结直肠癌细胞Caco-2细胞周期、迁移、侵袭和凋亡的影响及可能机制。方法RT-qPCR检测33例结直肠癌患者的癌组织和癌旁组织中SLC16A1-AS1和miR-450b-5p的表达水平。体外培养Caco-2细胞,双荧光素酶报告基因实验验证SLC16A1-AS1和miR-450b-5p靶向关系。将Caco-2细胞分为pcDNA组、pcDNA-SLC16A1-AS1组、pcDNA-SLC16A1-AS1+miR-NC组、pcDNA-SLC16A1-AS1+miR-450b-5p mimics组,流式细胞术检测细胞周期和细胞凋亡,Transwell检测细胞迁移和侵袭,Western blot检测Bcl-2和Bax蛋白表达。结果与癌旁组织比较,结直肠癌组织中SLC16A1-AS1的表达降低(P<0.05),而miR-450b-5p的表达升高(P<0.05)。SLC16A1-AS1在Caco-2细胞中靶向负调控miR-450b-5p的表达。与pcDNA组比较,pcDNA-SLC16A1-AS1组Caco-2细胞的细胞周期G0-G1期延长(P<0.05),S期缩短(P<0.05),细胞迁移和侵袭数及Bcl-2蛋白表达降低(P<0.05),凋亡率和Bax蛋白表达升高(P<0.05)。与pcDNA-SLC16A1-AS1+miR-NC组比较,pcDNA-SLC16A1-AS1+miR-450b-5p mimic组Caco-2细胞的细胞周期G0-G1期缩短(P<0.05),S期延长(P<0.05),细胞迁移和侵袭数及Bcl-2蛋白表达升高(P<0.05),凋亡率和Bax蛋白表达降低(P<0.05)。结论SLC16A1-AS1在结直肠癌组织中表达呈低表达,过表达SLC16A1-AS1可靶向负调控miR-450b-5p抑制结直肠癌Caco-2细胞的细胞周期进程、迁移和侵袭,并促进Caco-2细胞凋亡。

Objective To investigate the effects of SLC16A1-AS1 on cell cycle,migration,invasion and apoptosis of colorectal cancer cell Caco-2,and to explore possible action mechanism.Methods RT-qPCR was used to detect the expression levels of SLC16A1-AS1 and miR-450b-5p in cancer tissues and adjacent tissues of 33 patients with colorectal cancer.Caco-2 cells were cultured in vitro,and the dual luciferase reporter gene experiment was used to verify the regulatory correlation between SLC16A1-AS1 and miR-450b-5p.And Caco-2 cells were divided into pcDNA group,pcDNA-SLC16A1-AS1 group,pcDNA-SLC16A1-AS1+miR-NC group and pcDNA-SLC16A1-AS1+miR-450b-5p group.Flow cytometry was used to detect cell cycle and apoptosis,Transwell was used to detect cell migration and invasion,and Western Blot was used to detect the expression levels of Bcl-2 and Bax protein.Results Compared with those in adjacent tissues,the expression levels of SLC16A1-AS1 in colorectal cancer tissue were significantly decreased(P<0.05),however,the expression levels of miR-450b-5p were significantly increased(P<0.05).SLC16A1-AS1 negatively regulated the expression of miR-450b-5p in Caco-2 cells.Compared with that in pcDNA group,the cell cycle G0-G1 phase in pcDNA-SLC16A1-AS1 group was significantly prolonged(P<0.05),but the S phase was significantly shortened(P<0.05),and the number of cell migration and invasion and the protein expression levels of Bcl-2 were significantly decreased(P<0.05),while the cell apoptosis rate and and the expression levels of Bax were significantly increased(P<0.05).Compared with that in pcDNA-SLC16A1-AS1+miR-NC group,the cell cycle G0-G1 phase of Caco-2 cells in pcDNA-SLC16A1-AS1+miR-450b-5p mimic was group significantly shortened(P<0.05),but the S phase was significantly prolonged(P<0.05),and the number of cell migration and invasion and the expression levels of Bcl-2 were significantly increased(P<0.05),while the cell apoptosis rate and expression levels of Bax were significantly decreased(P<0.05).Conclusion The expression levels of SLC16A1-AS1 are decreased in colorectal cancer tissues,and overexpression of SLC16A1-AS1 can inhibit the cell cycle progression,migration and invasion of colorectal cancer Caco-2 cells and can promote cell apoptosis by negatively regulating miR-450b-5p.