卵巢癌ceRNA网络及预后模型的构建与验证

Construction and validation of ceRNA network and prognosis model for ovarian cancer

  • 摘要: 目的:基于卵巢癌竞争性内源RNA(ceRNA)调控网络构建预后风险模型,为预测卵巢癌患者预后规律及优化诊疗方案提供参考依据。方法:从Gene Expression Omnibus database(GEO)数据库中下载卵巢癌基因表达数据集,运用limma软件包分析筛选卵巢癌差异表达基因并构建lncRNA-microRNA-mRNA(ceRNA)调控网络。选取两个含有患者临床预后信息的数据集分别作为训练集和验证集构建卵巢癌预后风险模型,首先利用单因素Cox 回归筛选出调控网络中与预后相关的基因,其次运用多因素Cox回归构建疾病预后风险模型。再次采用生存曲线(K-M 曲线)和受试者工作特征曲线(ROC曲线)等方式评价模型的效能和稳健性。最后通过实时荧光定量聚合酶链式反应(RT-qPCR)验证模型基因在临床样本的表达水平。结果:筛选得到733 个卵巢癌差异表达基因,通过预测差异基因之间的调控关系成功构建包含134 个差异基因的ceRNA 调控网络。运用Cox回归构建得到包含5个关键基因(WASF3SNAI2PDE8BPCDH17RNF128)的卵巢癌多基因预后模型,K-M 曲线提示高风险组较低风险组的总体生存率更好(P< 0.01),训练集第1、第2、第3 年ROC 曲线下面积分别为0.702、0.737、0.741,验证第1、第2、第3年ROC曲线下面积分别为0.652、0.653、0.667,证实模型具有良好的预测效能和稳健性。基因表达验证表明WASF3SNAI2PDE8B 表达在卵巢癌组织中下调并且差异具有统计学意义(P< 0.05),PCDH17RNF128 呈下调趋势。结论:基于ceRNA调控网络所构建的预后风险模型在卵巢癌预后诊断中具有良好的效能和稳健性,有利于进一步指导临床治疗。

     

    Abstract: Objective:To construct a prognosis risk model model based on the competitive endogenous RNA(ceRNA)regulatory network of ovarian cancer to provide a reference for predicting the prognosis of ovarian can-cer patients and optimizing diagnosis and treatment plans.Methods:Ovarian cancer gene expression datasets were downloaded from the Gene Expression Omnibus database (GEO), “limma”software package was used to analyze and screen the differentially expressed genes of ovarian cancer, and the lncRNA-microRNA-mRNA(ceR-NA) regulatory network was constructed.Two datasets containing clinical prognosis information of patients were selected as the training set and the verification set to construct a prognosis risk model model of ovarian cancer.Firstly, univariate Cox regression analysis was utilized to screen out prognostic genes in the regulatory network, and secondly, the multivariate Cox regression analysis was used to construct prognosis risk model of ovarian can-cer.Furthermore, the survival curve (K-M curve) and receiver operating characteristic curve (ROC curve) were used to evaluate the effectiveness and robustness of the model.Finally, real-time fluorescence quantitative poly-merase chain reaction (RT-qPCR) was used to verify the expression level of the model genes in clinical samples.Results:733 differentially expressed genes in ovarian cancer were screened, and a ceRNA regulatory network containing 134 differentially expressed genes was suc-cessfully constructed by predicting the regulatory rela-tionships among the differentially expressed genes.A multi-gene prognostic model of ovarian cancer con-taining five key genes(WASF3, SNAI2, PDE8B, PCDH17, RNF128)was constructed by Cox regression.The KM curve showed that the overall survival rate of the high-risk group was better than that of the lower risk group(P< 0.01), the areas under the ROC curve in the 1st, 2nd and 3rd years of the training set were 0.702, 0.737 and 0.741, respectively, and the areas under the ROC curve in the 1st, 2nd and 3rd years of the validation set were 0.652, 0.653 and 0.667, respectively, confirming that the model had good predictive efficiency and robustness.Gene expression verification showed that WASF3, SNAI2 and PDE8B expressions were down-regulated in ovari-an cancer tissues and the differences were statistically significant(P< 0.05), and PCDH17 and RNF128 showed a down-regulation trend.Conclusion:The prognosis risk model constructed based on ceRNA regulatory network has good efficacy and robustness in the prognosis diagnosis of ovarian cancer, which is conducive to further guid-ing clinical treatment.

     

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