子宫内膜癌ceRNA的构建及LINC-FAM138B对HEC1B细胞恶性生物学行为的影响

Construction of endometrial carcinoma ceRNA and the effect of LINC-FAM138B on the malignant biological behavior of HEC1B cells

目的:构建子宫内膜癌(endometrial carcinoma,EC)竞争性内源RNA(competing endogenous RNA,ceRNA)并研究LINC-FAM138B对HEC1B细胞恶性生物学行为的影响。方法:从TCGA数据库下载EC患者的表达谱,筛选差异基因并构建ceRNA网络。STRING结合Cytoscape进行蛋白互作分析及筛选Top10基因,筛选与预后相关的基因,RT-qPCR实验验证LINC-FAM138B、hsa-mir-429和PRKG1在EC患者的癌组织及癌旁组织样本(n=7)中的表达水平。将细胞分为OV LINC-FAM138B组、Sh LINC-FAM138B组、OV NC组和Sh NC组,通过CCK-8实验检测细胞增殖能力;划痕实验检测细胞迁移能力;Transwell实验检测细胞侵袭能力;流式细胞术检测细胞凋亡能力。结果:筛选到457、65和1119个差异lncRNA、miRNA和mRNA。获得具有823个调控关系对的ceRNA网络。结合Top10基因和生存分析结果发现PRKG1是LINC-FAM138B结合hsa-mir-429靶向基因中得分最高的基因。PRKG1、RBFOX3、RGS2、SLC2A4高表达的EC患者预后显著比低表达的患者差(P<0.05)。RT-qPCR实验结果表明,与癌旁组织相比,LINC-FAM138B和PRKG1在EC癌组织中低表达(P<0.05),而hsa-mir-429在EC癌组织中高表达(P<0.05)。与OV NC组比较,OV LINC-FAM138B组中HEC1B细胞的增殖、迁移、侵袭能力显著下降(P<0.05),凋亡能力显著上升(P<0.01)。与Sh NC组比较,Sh LINC-FAM138B组中HEC1B细胞的增殖、迁移、侵袭能力显著上升(P<0.01),凋亡能力显著下降(P<0.05)。结论:构建了EC的ceRNA网络,且发现LINC-FAM138B抑制HEC1B细胞的增殖、迁移和侵袭及促进细胞的凋亡。

Objective:To construct competing endogenous RNA(ceRNA)from endometrial carcinoma(EC)and study the effect of LINC-FAM138B on the malignant biological behavior of HEC1B cells.Methods:Download the expression profile of EC patients from the TCGA database and screen for differential genes.Analyze differential genes and construct a ceRNA network.STRING combined with the Cytoscape database to perform protein interaction analysis and screen the Top10 gene,and screen key genes significantly related to prognosis.RT-qPCR experiments verified the expression levels of LINC-FAM138B,hsa-mir-429,and PRKG1 in cancer and adjacent tissues of EC patients(n=7).The cells were divided into four groups:OV LINC-FAM138B,Sh LINC-FAM138B,OV NC,Sh NC.The proliferation ability of the cells was detected by CCK-8 assay.Scratch test was used to detect the migration ability of cells,and Transwell assay was used to detect the invasion ability of cells.The ability of apoptosis was detected by flow cytometry.Results:457,65 and 1119 differentially expressed lncRNAs,miRNAs,and mRNAs were screened.A ceRNA network with 823 regulatory relationship pairs was obtained.Combining the Top10 gene and survival analysis results,it was found that PRKG1 had the highest score among LINC-FAM138B binding hsa-mir-429 targeted genes.In survival analysis,the prognosis of EC patients with high expression of PRKG1,RBFOX3,RGS2,and SLC2A4 was significantly worse than those with low expression(P<0.05).The results of RT-qPCR experiments showed that LINC-FAM138B and PRKG1 were lower expressed in EC cancer tissues than in adjacent tissues ( P <0.05),while hsa-mir-429 was higher expressed in EC cancer tissues ( P <0.05).Compared with OV NC group,the proliferation,migration,and invasion ability of HEC1B cells in OV LINC-FAM138B group decreased significantly ( P <0.05),while the apoptosis ability increased significantly ( P <0.01).Compared with the Sh NC group,the proliferation,migration,and invasion ability of HEC1B cells in the Sh LINC-FAM138B group significantly increased ( P <0.01),while the apoptosis ability significantly decreased ( P <0.05). Conclusion: We constructed a ceRNA network for EC and found that LINC-FAM138B inhibits the proliferation,migration,invasion,and promotes the apoptosis of HEC1B cells.