lncRNA PEG10 作为新的喉咽癌标志物及其调控机制研究

Survival rate for patients with early-stage hopharyngeal carcinoma is higher than that for those with late-stage carcinoma obviously. There is an important significance to diagnose hopharyngeal carcinoma early and mini-invasively in clinical. Tumor cells can release exosomes that contain lncRNAs into the blood. Investigation of aberrant circulating exosomal lncRNAs may provide new opportunity to discover tumor-related biomarkers. In our previous studies, high-throughput lncRNAs chip analysis was performed in tissue samples of hopharyngeal Survival rate for patients with early-stage hopharyngeal carcinoma is higher than that for those with late-stage carcinoma obviously. There is an important significance to diagnose hopharyngeal carcinoma early and mini-invasively in clinical. Tumor cells can release exosomes that contain lncRNAs into the blood. Investigation of aberrant circulating exosomal lncRNAs may provide new opportunity to find tumor-related biomarkers. In our previous studies, we found that the expression of lncRNA PEG10 was high in both tumor tissue and sera exosomes, which was related to the courses of EMT closely. The relationship between lncRNA PEG10 and tumorigenesis is not yet clear. Whether it can be used as a molecular marker for early diagnosis of tumors has not been studied. In this study, we intend to investigate the role of lncRNA PEG10 on the occurrence and development of hopharyngeal carcinoma by using cell EMT model and the vivo nude mice experiments. At the same time, we will further assess the diagnostic values of lncRNA PEG10 by RT-qPCR in large serum samples. It will be helpful to definite the function of lncRNA PEG10 in the course of occurrence and development of cancer, providing new ideas for the study of the pathogenesis and new targets for clinical diagnosis.

喉咽癌早期患者生存率明显高于晚期患者，有效微创性的早期诊断在临床上意义重大。肿瘤细胞能分泌大量含有lncRNA的外泌体到血循环中，检测异常lncRNA分子为肿瘤早期诊断提供了新机遇。我们前期研究发现lncRNA PEG10在喉咽癌组织及血清外泌体中均高表达，并与上皮间质细胞转化（EMT）过程关系密切。但lncRNA PEG10与喉咽癌发生的关系以及是否可作为分子标志物用于肿瘤早期诊断尚无研究。本课题拟通过建立细胞EMT模型、小鼠成瘤模型，研究lncRNA PEG10在喉咽癌发生发展过程中的作用机制，同时运用qRT-PCR技术对喉咽癌、癌前病变以及健康对照者血清进行大样本、多层次验证，评价lncRNA PEG10作为分子标记物应用于喉咽癌早期诊断价值。本研究将有助于明确 lncRNA PEG10在喉咽癌发生发展中的作用，也将为喉咽癌的实验室诊断提供依据。

本课题通过建立细胞EMT模型、小鼠成瘤模型，研究LncRNA PEG10在喉咽癌发生发展过程中的作用机制，同时运用qRT-PCR技术对喉咽癌、癌前病变以及健康对照者血清进行大样本、多层次验证，评价LncRNA PEG10作为分子标记物应用于喉咽癌早期诊断价值。研究结果如下：LncRNA PEG10参与TGF-β1诱导的NOK细胞，DOK细胞及Fuda 细胞EMT过程,其机制主要是通过靶向作用于PI3Kp85,减弱了Akt/Snail信号转导从而抑制上皮细胞发生EMT。此外我们也发现LncRNAPEG10作为分子"海绵"可吸附miR-21,上调miR-21的表达,从而进一步影响下游PI3Kp85/Akt/Snail信号通路,进而导致EMT。2. 4-NQO诱导小鼠喉咽黏膜由正常-癌前病变-癌”过程中，LncRNAPEG10在咽部黏膜组织中显著升高, Exosomal LncRNAPEG10表达水平在血清中亦显著升高。3.LncRNA PEG10表达水平在喉咽癌患者肿瘤组织中显著升高,并且与患者肿瘤恶性程度和不良预后有关。LncRNAPEG10可以作为喉咽癌预后的一项潜在的肿瘤标志物。4.Exosomal LncRNAPEG10表达水平在喉咽癌患者血清中显著升高,并且与患者恶性临床特征和不良预后有关。Exosomal LncRNAPEG10可以作为喉咽癌早期诊断、筛查及预后的一项潜在的肿瘤标志物。