circAGFG1/miR-487a-3p轴调控卵巢癌SKOV3细胞增殖、迁移、侵袭及EMT的实验研究

Experimental study of circAGFG1/miR-487a-3p axis regulating the proliferation, migration,invasion and EMT of ovarian cancer SKOV3 cells

目的 探讨circAGFG1对卵巢癌SKOV3细胞增殖、迁移、侵袭及上皮间质转化(EMT)的影响和作用机制。方法 RT-qPCR检测31例卵巢癌患者的癌组织和对应的癌旁组织中circAGFG1和miR-487a-3p表达。体外培养SKOV3细胞,分别转染circAGFG1小干扰RNA、miR-487a-3p模拟物或抑制剂后,CCK-8法检测细胞增殖,Transwell法对细胞迁移和侵袭进行检测,Western blot对细胞中Ki-67、MMP-2、MMP-9、Vimentin和E-cadherin蛋白表达进行检测。双荧光素酶报告基因实验验证circAGFG1和miR-487a-3p调控关系。结果 circAGFG1在卵巢癌组织中表达升高,miR-487a-3p表达降低。下调circAGFG1或上调miR-487a-3p阻碍SKOV3细胞增殖、迁移、侵袭及EMT。circAGFG1在SKOV3细胞中靶向负调控miR-487a-3p表达。敲减miR-487a-3p逆转下调circAGFG1对SKOV3细胞增殖、迁移、侵袭及EMT的影响。结论 下调circAGFG1可能通过靶向上调miR-487a-3p抑制卵巢癌细胞增殖、迁移和、侵袭及EMT。

Objective To investigate the effect and mechanism of circAGFG1 on the proliferation, migration, invasion and epithelial-mesenchymal transition(EMT) of ovarian cancer SKOV3 cells. Methods RT-qPCR was used to detect the expression levels of circAGFG1 and miR-487a-3p in cancer tissues and corresponding adjacent tissues of 31 ovarian cancer patients. SKOV3 cells were cultured in vitro and transfected circAGFG1 small interfering RNA, miR-487a-3p mimic or inhibitor. CCK-8 method was used to detect cell proliferation. Cell migration and invasion were detected by Transwell. Western blot was used to detect the expression of Ki-67, MMP-2, MMP-9, Vimentin and E-cadherin protein in cells. The dual luciferase reporter gene experiment verified the regulatory relationship between circAGFG1 and miR-487a-3p. Results The expression of circAGFG1 was increased in ovarian cancer tissues, but the expression of miR-487a-3p was decreased. Down-regulating circAGFG1 or up-regulating miR-487a-3p hindered proliferation, migration, invasion and EMT of SKOV3 cells. circAGFG1negatively regulated the expression of miR-487a-3p in SKOV3 cells. Knockdown of miR-487a-3p reversed the effect of down-regulating circAGFG1 on the proliferation, migration, invasion and EMT of SKOV3 cells. Conclusion Down-regulation of circAGFG1 may inhibit the proliferation, migration, invasion and EMT of ovarian cancer cells by targeting to up-regulate miR-487a-3p.