甘草素调控circ_0006916/microRNA-513a-3p对结直肠癌细胞LoVo生物行为的影响

The effect of liquiritigenin on the biological behavior of colorectal cancer cell LoVo by regulating circ_0006916/microRNA-513a-3p

目的:探讨甘草素对结直肠癌细胞LoVo生物行为的影响及其可能的作用机制。方法:用不同浓度的甘草素处理LoVo细胞,用脂质体转染将si-NC、si-circ_0006916分别转染至LoVo细胞,将pcDNA-circ_0006916转染至LoVo细胞后加入0.4 mmol/L甘草素处理24 h进行实验;MTT法、平板克隆形成实验分别检测细胞增殖及集落形成数;划痕实验与Transwell实验检测细胞迁移能力;流式细胞术检测细胞凋亡率;qRT-PCR法检测circ_0006916、微小RNA-513a-3p(miR-513a-3p)的表达量;双荧光素酶报告实验检测circ_0006916与miR-513a-3p的靶向关系;Western blot法检测Cleaved-caspase3蛋白表达量。结果:甘草素处理LoVo细胞后,细胞增殖抑制率、凋亡率、Cleaved-caspase3蛋白水平和miR-513a-3p的表达量升高(P<0.05),而集落形成数和迁移细胞数减少(P<0.05),circ_0006916的表达量和划痕愈合率降低(P<0.05),且呈剂量依赖性;circ_0006916与miR-513a-3p存在靶向调控作用;转染si-circ_0006916可促进miR-513a-3p表达(P<0.05),细胞增殖抑制率、凋亡率和Cleaved-caspase3蛋白水平升高(P<0.05),集落形成数和迁移细胞数减少(P<0.05),划痕愈合率降低(P<0.05);转染pcDNA-circ_0006916可降低甘草素对LoVo细胞生物行为的作用。结论:甘草素可通过调控circ_0006916/miR-513a-3p表达而抑制结直肠癌细胞增殖、克隆形成及迁移,并可促进细胞凋亡。

Objective:To explore the effect of liquiritigenin on the biological behavior of colorectal cancer cells LoVo and its possible mechanism.Methods:Different concentrations of liquiritigenin were used to treat LoVo cells.The si-NC and si-circ_0006916 were transfected into LoVo cells by liposome transfection.After pcDNA-circ_0006916 was transfected into LoVo cells,0.4 mmol/L liquiritigenin was added for 24 hours to perform a response experiment.MTT method and plate colony formation experiment were used to detect cell proliferation and colony formation.Scratch test and Transwell test to detect cell migration ability.Flow cytometry to detect cell apoptosis rate.qRT-PCR method was used to detect the expression of circ_0006916 and microRNA-513 a-3 p(miR-513 a-3 p).The dual luciferase reporter experiment was used to detect the targeting relationship between circ_0006916 and miR-513 a-3 p.Western blot method was used to detect the expression of Cleaved-caspase3 protein.Results:After liquiritigenin treatment of LoVo cells,the cell proliferation inhibition rate,apoptosis rate,the protein level of Cleaved-caspase3 and the expression of miR-513 a-3 p were increased(P<0.05),while the number of colonies formed and the number of migrating cells were decreased(P<0.05),the expression level of circ_0006916 and the scratch healing rate were decreased(P<0.05),and were dose-dependent.circ_0006916 and miR-513 a-3 p had a targeted regulation effect.Transfection of si-circ_0006916 could promote the expression of miR-513 a-3 p(P<0.05),and could increase the cell proliferation inhibition rate,apoptosis rate and the protein level of Cleaved-caspase3(P<0.05),and could decrease the number of colonies formed and the number of migrating cells(P<0.05),the healing rate of scratches was reduced(P<0.05).Transfection of pcDNA-circ_0006916 could reduce the effect of liquiritigenin on the biological behavior of LoVo cells.Conclusion:Liquiritigenin could inhibit the proliferation,clone formation and migration of colorectal cancer cells by regulating the expression of circ_0006916/miR-513 a-3 p,and can promote cell apoptosis.