LINC01128在子宫内膜癌组织中的表达及其调节miR-367-3p/KLF5轴对细胞恶性进展的影响

Expression of LINC01128 in Endometrial Cancer Tissue and Its Effect on Cell Malignant Progression by Regulating miR-367-3p/KLF5 Axis

该文旨在探究LINC01128在子宫内膜癌(EC)组织中的表达情况及其调节miR-367-3p/Krüppel样因子5(KLF5)轴对细胞恶性进展的影响。qRT-PCR检测LINC01128、miR-367-3p和KLF5 mRNA在EC组织、EC细胞系HEC-1A、人子宫内膜上皮细胞CP-H058中的表达水平。将体外培养的HEC-1A细胞随机分为:NC组(正常培养)、si-LINC01128组、si-NC组、miR-367-3p mimic组及miR-NC组。si-NC组、si-LINC01128组、miR-367-3p mimic组、miR-NC组分别转染si-NC、si-LINC01128、miR-367-3pmimic、miR-NC。CCK-8法、TUNEL染色分别检测HEC-1A细胞增殖、凋亡情况;Transwell、划痕实验分别检测HEC-1A细胞侵袭、迁移情况;双荧光素酶报告基因实验验证LINC01128与miR-367-3p、miR-367-3p与KLF5的靶向关系;Western blot检测细胞中KLF5表达情况。与癌旁组织相比,EC组织中LINC01128和KLF5 mRNA表达水平升高,miR-367-3p表达水平降低;与CP-H058细胞相比,EC细胞HEC-1A中LINC01128和KLF5 mRNA表达水平升高,miR-367-3p表达水平降低(P<0.05)。NC组与si-NC组LINC01128、miR-367-3p、KLF5mRNA表达水平以及HEC-1A细胞增殖、凋亡、迁移、侵袭情况无显著差异(P>0.05)。与si-NC组相比,siLINC01128组LINC01128、KLF5 mRNA表达水平以及D450值、迁移率、侵袭数、Bcl-2表达水平均降低,miR-367-3p表达水平、凋亡率及Bax表达水平均升高(P<0.05)。NC组与miR-NC组LINC01128、miR-367-3p、KLF5mRNA表达水平以及HEC-1A细胞增殖、凋亡、迁移及侵袭情况无显著差异(P>0.05)。与miR-NC组对比,miR-367-3p mimic组LINC01128表达水平无显著差异(P>0.05),KLF5mRNA表达水平、D450值、迁移率、侵袭数及Bcl-2表达水平均降低,miR-367-3p表达水平、凋亡率及Bax表达水平均升高(P<0.05)。双荧光素酶报告基因实验结果显示,miR-367-3p和LINC01128、KLF5均存在靶向调控关系。LINC01128在EC组织和细胞中呈高表达,干扰LINC01128可通过调节miR-367-3p/KLF5轴抑制EC恶性进展。

This study aims to investigate the expression of LINC01128 in EC(endometrial cancer)tissue and its effect on cell malignant progression by regulating the miR-367-3p/KLF5(Krüppel like factor 5)axis.qRT-PCR was applied to detect the expression levels of LINC01128,miR-367-3p,and KLF5 mRNA in EC tissue,cell line HEC-1A,human endometrial epithelial cells CP-H058.HEC-1A cells cultured in vitro were randomly separated into five groups:NC group(normal culture),si-LINC01128 group,si-NC group,miR-367-3p mimic group,and miR-NC group.And the si-NC group,si-LINC01128 group,miR-367-3p mimic group,and miR-NC group were transfected with si-NC,si-LINC01128,miR367-3p mimic,and miR-NC,respectively.CCK-8 method and TUNEL staining were applied to detect the proliferation and apoptosis of HEC-1A cells,respectively.Transwell and scratch experiments were applied to detect the invasion and migration of HEC-1A cells,respectively.Dual luciferase reporter gene experiment was applied to verify the targeting relationship between LINC01128 and miR-367-3p,and between miR-367-3p and KLF5.Western blot was applied to detect the expression of KLF5 in cells.Compared with adjacent tissues,the expression of LINC01128 and KLF5 mRNA in EC tissues was increased,while miR-367-3p expression was decreased;compared with CP-H058 cell,the expression of LINC01128 and KLF5 mRNA in EC cell HEC-1A was increased,while miR-367-3p expression was reduced(P<0.05).There was no significant difference in the expression of LINC01128,miR-367-3p,KLF5 mRNA,as well as HEC-1A cell proliferation,apoptosis,migration,and invasion between the NC group and the si-NC group(P>0.05).Compared with the si-NC group,the expression of LINC01128 and KLF5 mRNA,D450 value,migration rate,invasion number,and Bcl-2 expression in the si-LINC01128 group were all reduced,while miR-367-3p expression,apoptosis rate,and Bax expression were all increased(P<0.05).There was no great difference in the expression of LINC01128,miR-367-3p,KLF5 mRNA,as well as HEC-1A cell proliferation,apoptosis,migration,and invasion between the NC group and the miR-NC group(P>0.05).Compared with the miR-NC group,there was no great difference in LINC01128 expression in the miR-367-3p mimic group(P>0.05),the KLF5 mRNA expression,D450 value,migration rate,invasion number,and Bcl-2 expression were all reduced,while miR-367-3p expression,apoptosis rate,and Bax expression were all increased(P<0.05).The results of dual luciferase reporter gene experiment showed that miR-367-3p had targeted regulatory relationship with LINC01128 and KLF5.LINC01128 is highly expressed in EC tissues and cells,and interference with LINC01128 can inhibit malignant progression of EC by regulating the miR-367-3p/KLF5 axis.