人口腔鳞状细胞癌发病节点lncRNA的筛选、验证及意义

The screening and verification of human squamous cell carcinoma pathogenesis node lncRNA and its clinical significance

目的应用微阵列芯片技术检测长链非编码RNA(long non-codingRNA,lncRNA)在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)组织中的表达谱,筛选OSCC发病过程中节点lncRNA,研究其表达和临床意义。方法利用lncRNA表达谱芯片,对3例OSCC及对应正常组织的转录本进行高通量筛查,确定OSCC中的lncRNAs和mRNAs差异表达谱。利用lncRNA与mRNA特异性表达的标准化信号强度,构建差异基因共表达网络,挖掘节点基因,分析其表达水平与临床病理特征及患者预后之间的关系。选取部分差异表达的lncRNA,借助qRT-PCR技术对芯片结果进行验证。结果实验共筛选出差异表达lncRNAs2 294条和mRNAs 1 938条。LncRNA TMPRSS11BNL、lnc-WRN-10:1是共表达网络的节点基因,在OSCC组织中低表达,并与患者临床分期、淋巴结转移、远处转移和生存状态有关,低表达lncRNA TMPRSS11BNL、lnc-WRN-10:1 OSCC患者有着较短的无进展生存时间和总生存时间。qRT-PCR验证结果与芯片结果相一致。结论LncRNA TMPRSS11BNL、lnc-WRN-10:1有望成为成为OSCC早期诊断、治疗及预后评估的分子标志物。

Objective To assess the expression profile of long non-coding RNA(lncRNA)in oral squamous cell carcinoma(OSCC)tissues by microarray technique,and to screen the node lncRNAs,in the pathogenesis of OSCC and to study its expression and clinical significance.Methods High-throughput screening of transcripts from 3 cases of OSCC and corresponding normal tissues was performed by using lncRNA microarray to determine the differential expression profiles of lncRNAs and mRNAs in OSCC.The differential gene co-expression network was constructed by using the standardized signal intensity of specific expression of lncRNA and mRNA to excavate node gene.The correlations between the expression level of node genes and the clinicopathological features and prognosis of the patients were analyzed.Partial differentially expressed lncRNA,was selected to verify the chip results by qRT-PCR technique.Results A total of 2 294 differentially expressed lncRNAs and 1 938 mRNAs genes were screened.LncRNA TMPRSS11BNL,lnc-WRN-10:1 was the node gene of the co-expression network.The low expression of lncRNA TMPRSS11BNL and lnc-WRN-10:1 in OSCC tissues was associated with clinical stage,lymph node metastasis,distant metastasis and survival status.OSCC patients with low expression of lncRNA TMPRSS11BNL and lnc-WRN-10:1 had shorter progression-free survival and overall survival.The results of qRT-PCR verification were consistent with the results of microarray.Conclusion LncRNA TMPRSS11BNL and lnc-WRN-10:1 may be molecular markers for early diagnosis,treatment and prognosis of OSCC.