FOXD2-AS1、MIR4435-1HG和HOXC-AS2联合诊断胃癌的临床价值及机制研究

Diagnostic value of combined detection of FOXD2-AS1,MIR4435-1HG and HOXC-AS2 and mechanism of the three genes in gastric cancer

目的:筛选胃癌中差异性表达的长链非编码RNA(lncRNA),探讨代表性lncRNA在胃癌诊断中的潜在价值及相关机制。方法:从TCGA数据库中筛选在胃癌中差异性表达的lncRNA,通过GSE179252和GSE184336数据集以及qRT-PCR检测胃癌患者血清中lncRNA的表达水平对结果进行验证。绘制ROC曲线评价代表性lncRNA单独及联合检测的诊断效能。分析HOXC-AS2与胃癌患者临床病理参数的相关性。利用在线数据库对HOXC-AS2进行下游靶基因预测以及PPI蛋白互作分析,构建HOXC-AS2-miRNA-mRNA调控网络。结果:从TCGA数据库中筛选出3个在胃癌中差异性表达的lncRNA。TCGA-STAD、GSE179252和GSE184336数据集中FOXD2-AS1、MIR4435-1HG和HOXC-AS2联合检测胃癌的ROC曲线下面积(AUC)分别为0.994、0.958和0.921。血清中检测结果发现与胃良性疾病患者相比,FOXD2-AS1、MIR4435-1HG和HOXC-AS2在胃癌患者血清中表达升高(P<0.001),三者联合检测诊断胃癌的AUC为0.843,灵敏度和特异度分别为80%和86%。HOXC-AS2与胃癌肿瘤分级有相关性(P<0.05)。HOXC-AS2可与下游mRNA竞争miR-9-5p等miRNA结合位点,调控DLX6与HOXC蛋白家族之间的相互作用。结论:FOXD2-AS1、MIR4435-1HG和HOXC-AS2的联合检测可用作胃癌的诊断标志物,HOXC-AS2可作为ceRNA促进胃癌进展。

Objective:To screen out the long non-coding RNAs(lncRNA)differentially expressed in gastric cancer,and investigate the clinical diagnostic value and mechanism of representative lncRNA.Methods:Differentially expressed lncRNA in gastric cancer were screened out from TCGA database,and verified using GSE184336 and GSE179252 datasets.qRT-PCR was performed to detect the candidate lncRNA in serum for further investigation.ROC curves were mapped to assess the diagnostic efficacy by detecting lncRNA alone or combined detection of the three genes.The correlation between HOXC-AS2 and clinicopathological parameters were analyzed.Online database was used to predict downstream miRNAs and analyze PPI protein interaction associated with HOXC-AS2.Then a HOXC-AS2-miRNA-mRNA network was constructed.Results:Three differentially expressed lncRNA were screened out in gastric cancer.ROC curve analysis showed that FOXD2-AS1,MIR4435-1HG and HOXC-AS2 had high value for combined diagnosis of gastric cancer in TCGA-STAD,GSE179252 and GSE184336 datasets(AUC=0.994,0.958 and 0.921,respectively).Compared with gastric tissues from patients with benign neoplasm,the expression levels of serum FOXD2-AS1,MIR4435-1HG and HOXC-AS2 were higher in gastric cancer patients(all P<0.001).The AUC of combined diagnosis of gastric cancer was 0.843,and the sensitivity and specificity were 80%and 86%,respectively.The level of HOXC-AS2 expression was correlated with tumor grade(P<0.05).HOXC-AS2 could regulate the interaction between DLX6 and HOXC protein family by binding with downstream miRNAs,such as miR-9-5p.Conclusion:Combined detection of FOXD2-AS1,MIR4435-1HG and HOXC-AS2 can be used markers for gastric cancer,and HOXC-AS2 may be role of ceRNA in promoting progression of gastric cancer.