干扰LINC00466通过miR-493-3p/MIF抑制子宫内膜癌RL95-2细胞恶性生物学行为

Interference with LINC00466 inhibits the malignant biological behavior of endometrial cancer RL95-2 cells through miR-493-3p/MIF

目的探讨LINC00466对子宫内膜癌RL95-2细胞生物学行为的影响及其作用机制。方法收集43例子宫内膜癌患者癌组织和癌旁组织,qRT-PCR法检测组织中LINC00466和miR-493-3p表达。体外培养RL95-2细胞,转染LINC00466小干扰RNA或miR-493-3p模拟物、或共转染LINC00466小干扰RNA与miR-493-3p抑制剂后,CCK-8法、Transwell实验分别检测细胞增殖抑制率、迁移和侵袭,qRT-PCR和Western blot分别检测细胞中巨噬细胞移动抑制因子(MIF)mRNA和蛋白表达。双荧光素酶报告基因实验对LINC00466和miR-493-3p调控关系进行验证。两组间比较采用t检验。结果子宫内膜癌组织中LINC00466的表达量较癌旁组织(2.66±0.34比0.95±0.21)升高,miR-493-3p的表达量较癌旁组织(0.42±0.10比0.98±0.15)降低,差异有统计学意义(P均<0.001)。干扰LINC00466或过表达miR-493-3p增加RL95-2细胞增殖抑制率,减少细胞迁移数、侵袭数及细胞中MIF mRNA和蛋白表达(P<0.001)。LINC00466靶向结合miR-493-3p,且干扰LINC00466的RL95-2细胞中miR-493-3p表达增加。敲减miR-493-3p逆转干扰LINC00466对RL95-2细胞增殖、迁移和侵袭及MIF表达的影响。结论LINC00466在子宫内膜癌组织中表达上调,干扰LINC00466可能通过靶向上调miR-493-3p和阻碍MIF表达来抑制子宫内膜癌RL95-2细胞增殖、迁移和侵袭。

Objective To investigate the effect of LINC00466 on the biological behavior of endometrial cancer RL95-2 cells and its mechanism.Methods The cancer tissues and adjacent tissues of 43 patients with endometrial cancer were collected,and the expression of LINC00466 and miR-493-3p in the tissues were detected by qRT-PCR.RL95-2 cells were cultured in vitro and transfected with LINC00466 small interfering RNA or miR-493-3p mimic,or co-transfected with LINC00466 small interfering RNA and miR-493-3p inhibitor,and then CCK-8 method and Transwell assay were used to detect cell proliferation inhibition rate,migration and invasion.qRT-PCR and Western blot were used to detect the mRNA and protein expression of macrophage migration inhibitory factor(MIF)in cells,respectively.The regulatory relationship between LINC00466 and miR-493-3p was verified by the dual-luciferase reporter gene experiment.Comparison between the two groups was performed by t test.Results The expression of LINC00466 in endometrial cancer tissues was higher than that in adjacent tissues(2.66±0.34 vs 0.95±0.21),but the expression of miR-493-3p was lower than that in adjacent tissues(0.42±0.10 vs 0.98±0.15),and the differences were statistically significant(both P<0.001).Interfering with LINC00466 or overexpressing miR-493-3p increased the proliferation inhibition rate of RL95-2 cells,but decreased the number of cell migration and invasion and the mRNA and protein expression of MIF in cells(P<0.001).LINC00466 targets miR-493-3p,and the expression of miR-493-3p was increased in RL95-2 cells that interfered with LINC00466.Knockdown of miR-493-3p reversed the effects of interfering with LINC00466 on proliferation,migration,invasion and the expression of MIF in RL95-2 cells.Conclusion The expression of LINC00466 was up-regulated in endometrial cancer tissues.Interfering with LINC00466 may inhibit the proliferation,migration and invasion of endometrial cancer RL95-2 cells by targeting up-regulation of miR-493-3p and blocking MIF expression.