Abstract：
Mounting evidence suggests that noncoding RNAs (lncRNAs) were involved in various human cancers. However, the role of these lncRNAs in HPV-driven cervical cancer (CC) has not been extensively studied. Considering that HR-HPV infections contribute to cervical carcinogenesis by regulating the expression of lncRNAs, miRNAs and mRNAs, we aim to systematically analyze lncRNAs and mRNAs expression profile to identify novel lncRNAs-mRNAs co-expression networks and explore their potential impact on tumorigenesis in HPV-driven CC.
LncRNA/mRNA microarray technology was utilized to identify the differentially expressed lncRNAs (DElncRNAs) and mRNAs (DEmRNAs) in HPV-16 and HPV-18 cervical carcinogenesis compared to normal cervical tissues. Venn diagram and weighted gene co-expression network analysis (WGCNA) were used to identify the hub DElncRNAs/DEmRNAs that were both significantly correlated with HPV-16 and HPV-18 CC patients. LncRNA-mRNA correlation analysis and functional enrichment pathway analysis were performed on these key DElncRNAs/DEmRNAs in HPV-16 and HPV-18 CC patients to explore their mutual mechanism in HPV-driven CC. A lncRNA-mRNA co-expression score (CES) model was established and validated by using the Cox regression method. Afterward, the clinicopathological characteristics were analyzed between CES-high and CES-low groups. In vitro, functional experiments were performed to evaluate the role of LINC00511 and PGK1 in cell proliferation, migration and invasion in CC cells. To understand whether LINC00511 play as an oncogenic role partially via modulating the expression of PGK1, rescue assays were used.
We identified 81 lncRNAs and 211 mRNAs that were commonly differentially expressed in HPV-16 and HPV-18 CC tissues compared to normal tissues. The results of lncRNA-mRNA correlation analysis and functional enrichment pathway analysis showed that the LINC00511-PGK1 co-expression network may make an important contribution to HPV-mediated tumorigenesis and be closely associated with metabolism-related mechanisms. Combined with clinical survival data, the prognostic lncRNA-mRNA co-expression score (CES) model based on LINC00511 and PGK1 could precisely predict patients\' overall survival (OS). CES-high patients had a worse prognosis than CES-low patients and the enriched pathways and potential targets of applicable drugs were explored in CES-high patients. In vitro experiments confirmed the oncogenic functions of LINC00511 and PGK1 in the progression of CC, and revealed that LINC00511 functions in an oncogenic role in CC cells partially via modulating the expression of PGK1.
Together, these data identify co-expression modules that provide valuable information to understand the pathogenesis of HPV-mediated tumorigenesis, which highlights the pivotal function of the LINC00511-PGK1 co-expression network in cervical carcinogenesis. Furthermore, our CES model has a reliable predicting ability that could stratify CC patients into low- and high-risk groups of poor survival. This study provides a bioinformatics method to screen prognostic biomarkers which leads to lncRNA-mRNA co-expression network identification and construction for patients\' survival prediction and potential drug applications in other cancers.
LncRNA/mRNA microarray technology was utilized to identify the differentially expressed lncRNAs (DElncRNAs) and mRNAs (DEmRNAs) in HPV-16 and HPV-18 cervical carcinogenesis compared to normal cervical tissues. Venn diagram and weighted gene co-expression network analysis (WGCNA) were used to identify the hub DElncRNAs/DEmRNAs that were both significantly correlated with HPV-16 and HPV-18 CC patients. LncRNA-mRNA correlation analysis and functional enrichment pathway analysis were performed on these key DElncRNAs/DEmRNAs in HPV-16 and HPV-18 CC patients to explore their mutual mechanism in HPV-driven CC. A lncRNA-mRNA co-expression score (CES) model was established and validated by using the Cox regression method. Afterward, the clinicopathological characteristics were analyzed between CES-high and CES-low groups. In vitro, functional experiments were performed to evaluate the role of LINC00511 and PGK1 in cell proliferation, migration and invasion in CC cells. To understand whether LINC00511 play as an oncogenic role partially via modulating the expression of PGK1, rescue assays were used.
We identified 81 lncRNAs and 211 mRNAs that were commonly differentially expressed in HPV-16 and HPV-18 CC tissues compared to normal tissues. The results of lncRNA-mRNA correlation analysis and functional enrichment pathway analysis showed that the LINC00511-PGK1 co-expression network may make an important contribution to HPV-mediated tumorigenesis and be closely associated with metabolism-related mechanisms. Combined with clinical survival data, the prognostic lncRNA-mRNA co-expression score (CES) model based on LINC00511 and PGK1 could precisely predict patients\' overall survival (OS). CES-high patients had a worse prognosis than CES-low patients and the enriched pathways and potential targets of applicable drugs were explored in CES-high patients. In vitro experiments confirmed the oncogenic functions of LINC00511 and PGK1 in the progression of CC, and revealed that LINC00511 functions in an oncogenic role in CC cells partially via modulating the expression of PGK1.
Together, these data identify co-expression modules that provide valuable information to understand the pathogenesis of HPV-mediated tumorigenesis, which highlights the pivotal function of the LINC00511-PGK1 co-expression network in cervical carcinogenesis. Furthermore, our CES model has a reliable predicting ability that could stratify CC patients into low- and high-risk groups of poor survival. This study provides a bioinformatics method to screen prognostic biomarkers which leads to lncRNA-mRNA co-expression network identification and construction for patients\' survival prediction and potential drug applications in other cancers.
摘要：
背景：越来越多的证据表明，非编码RNA（lncRNAs）与各种人类癌症有关。然而,这些lncRNAs在HPV驱动的宫颈癌(CC)中的作用尚未得到广泛研究.考虑到HR-HPV感染通过调节lncRNAs的表达促进宫颈癌的发生，miRNA和mRNA,我们旨在系统分析lncRNAs和mRNAs表达谱,以鉴定新的lncRNAs-mRNAs共表达网络,并探讨它们对HPV驱动的CC中肿瘤发生的潜在影响.
方法：利用LncRNA/mRNA微阵列技术来鉴定与正常宫颈组织相比，HPV-16和HPV-18宫颈癌发生中差异表达的lncRNA（DElncRNA）和mRNAs（DEmRNAs）。使用维恩图和加权基因共表达网络分析(WGCNA)来鉴定均与HPV-16和HPV-18CC患者显著相关的hubDElncRNAs/DEmRNAs。对HPV-16和HPV-18CC患者中的这些关键DElncRNA/DEmRNA进行LncRNA-mRNA相关性分析和功能富集通路分析，以探讨其在HPV驱动CC中的相互机制。建立lncRNA-mRNA共表达评分(CES)模型,采用Cox回归方法进行验证。之后,分析CES高组和CES低组的临床病理特征。体外，进行功能实验以评估LINC00511和PGK1在细胞增殖中的作用，CC细胞的迁移和侵袭。为了理解LINC00511是否部分地通过调节PGK1的表达而发挥致癌作用,使用了拯救测定。
结果：我们鉴定了与正常组织相比，在HPV-16和HPV-18CC组织中通常差异表达的81个lncRNAs和211个mRNAs。lncRNA-mRNA相关性分析和功能富集通路分析结果表明，LINC00511-PGK1共表达网络可能对HPV介导的肿瘤发生有重要贡献，并与代谢相关机制密切相关。结合临床生存数据，基于LINC00511和PGK1的预后lncRNA-mRNA共表达评分(CES)模型可以准确预测患者的总生存期(OS).高CES患者的预后比低CES患者差，并在高CES患者中探索了适用药物的富集途径和潜在靶标。体外实验证实了LINC00511和PGK1在CC进展中的致癌功能，并揭示LINC00511部分通过调节PGK1的表达在CC细胞中起致癌作用。
结论：一起，这些数据确定了共表达模块，这些模块为了解HPV介导的肿瘤发生的发病机理提供了有价值的信息，这突出了LINC00511-PGK1共表达网络在宫颈癌发生中的关键作用。此外，我们的CES模型具有可靠的预测能力,可以将CC患者分为低生存率和高生存率组.这项研究提供了一种生物信息学方法来筛选预后生物标志物，从而导致lncRNA-mRNA共表达网络的鉴定和构建，用于患者的生存预测和其他癌症的潜在药物应用。
方法：利用LncRNA/mRNA微阵列技术来鉴定与正常宫颈组织相比，HPV-16和HPV-18宫颈癌发生中差异表达的lncRNA（DElncRNA）和mRNAs（DEmRNAs）。使用维恩图和加权基因共表达网络分析(WGCNA)来鉴定均与HPV-16和HPV-18CC患者显著相关的hubDElncRNAs/DEmRNAs。对HPV-16和HPV-18CC患者中的这些关键DElncRNA/DEmRNA进行LncRNA-mRNA相关性分析和功能富集通路分析，以探讨其在HPV驱动CC中的相互机制。建立lncRNA-mRNA共表达评分(CES)模型,采用Cox回归方法进行验证。之后,分析CES高组和CES低组的临床病理特征。体外，进行功能实验以评估LINC00511和PGK1在细胞增殖中的作用，CC细胞的迁移和侵袭。为了理解LINC00511是否部分地通过调节PGK1的表达而发挥致癌作用,使用了拯救测定。
结果：我们鉴定了与正常组织相比，在HPV-16和HPV-18CC组织中通常差异表达的81个lncRNAs和211个mRNAs。lncRNA-mRNA相关性分析和功能富集通路分析结果表明，LINC00511-PGK1共表达网络可能对HPV介导的肿瘤发生有重要贡献，并与代谢相关机制密切相关。结合临床生存数据，基于LINC00511和PGK1的预后lncRNA-mRNA共表达评分(CES)模型可以准确预测患者的总生存期(OS).高CES患者的预后比低CES患者差，并在高CES患者中探索了适用药物的富集途径和潜在靶标。体外实验证实了LINC00511和PGK1在CC进展中的致癌功能，并揭示LINC00511部分通过调节PGK1的表达在CC细胞中起致癌作用。
结论：一起，这些数据确定了共表达模块，这些模块为了解HPV介导的肿瘤发生的发病机理提供了有价值的信息，这突出了LINC00511-PGK1共表达网络在宫颈癌发生中的关键作用。此外，我们的CES模型具有可靠的预测能力,可以将CC患者分为低生存率和高生存率组.这项研究提供了一种生物信息学方法来筛选预后生物标志物，从而导致lncRNA-mRNA共表达网络的鉴定和构建，用于患者的生存预测和其他癌症的潜在药物应用。
