circ_0016760调控miR-625对乳腺癌细胞生物行为的影响

Effects of circ_0016760 regulating miR-625 on the biological behavior of breast cancer cells in vitro

目的探讨circ_0016760对乳腺癌细胞生物行为的影响及其可能作用机制。方法qRT-PCR法检测乳腺癌组织与癌旁组织中circ_0016760、miR-625的表达量;体外培养人乳腺癌细胞T-47D,si-NC、si-circ_0016760、miR-NC、miR-625 mimics、anti-miR-NC、anti-miR-625、si-circ_0016760与anti-miR-625分别转染至T-47D细胞;双荧光素酶报告实验检测circ_0016760和miR-625的靶向关系;MTT法与平板克隆形成实验检测细胞增殖能力;划痕实验与Transwell小室实验检测细胞迁移能力;流式细胞术检测细胞凋亡率;Western blot法检测cleaved-caspase3蛋白表达量。结果乳腺癌组织中circ_0016760的表达量高于癌旁组织(P<0.05),miR-625的表达量低于癌旁组织(P<0.05);circ_0016760可靶向调节miR-625的表达;转染si-circ_0016760或转染miR-625 mimics后细胞活力和划痕愈合率降低(P<0.05),集落形成数和迁移细胞数减少(P<0.05),细胞凋亡率和cleaved-caspase3蛋白水平升高(P<0.05);转染anti-miR-625后细胞活力和划痕愈合率升高(P<0.05),集落形成数和迁移细胞数增多(P<0.05),细胞凋亡率和cleaved-caspase3蛋白水平降低(P<0.05);共转染si-circ_0016760与anti-miR-625能够逆转转染si-circ_0016760对T-47D细胞增殖、集落形成、迁移及凋亡的作用。结论干扰circ_0016760表达可通过上调miR-625表达而抑制乳腺癌细胞增殖、集落形成、迁移及促进细胞凋亡。

Objective To investigate the effects of circ_0016760 regulating miR-625 on the biological behavior of breast cancer cells in vitro,and to explore its possible action mechanism.Methods The qRT-PCR was used to detect the expression levels of circ_0016760 and miR-625 in breast cancer tissues and para-cancerous tissues.The si-NC,si-circ_0016760,miR-NC,miR-625 mimics,anti-miR-NC,anti-miR-625,si-circ_0016760 and anti-miR-625 were transferred respectively into in vitro cultured human breast cancer cells(T-47D).The dual luciferase reporter experiment was used to detect the targeting correlation between circ_0016760 and miR-625,and MTT and plate colony formation experiment were used to detect cell proliferation ability,and scratch test and Transwell chamber test were used to detect cell migration ability.Moreover flow cytometry detected the cell apoptosis rate,and Western Blot detected the expression levels of cleaved-caspase3 protein.Results The expression levels of circ_0016760 in breast cancer tissues were significantly higher than those in para-cancerous tissues(P<0.05),however,the expression levels of miR-625 were significantly lower than those in para-cancerous tissues(P<0.05).The circ_0016760 could targetedly regulate the expression of miR-625.After transfection with si-circ_0016760 or miR-625 mimics,the cell viability,scratch healing rate,the number of colony formation and the number of migrating cells were significantly decreased(P<0.05),while the apoptosis rate and the expression levels of cleaved-caspase3 protein were significantly increased(P<0.05).After transfection with anti-miR-625,the cell viability and scratch healing rate,and the number of colony formation and the number of migrating cells were significantly increased(P<0.05),while the apoptosis rate and the expression levels of cleaved-caspase3 protein were significantly decreased(P<0.05).Co-transfection of si-circ_0016760 and anti-miR-625 could reverse the effects of transfection of si-circ_0016760 on the proliferation,colony formation,migration and apoptosis of T-47D cells.Conclusion To interfere with the expression of circ_0016760 can inhibit the proliferation,colony formation,migration of breast cancer cell,and can promote cell apoptosis by up-regulating the expression of miR-625.