长链非编码RNA-Cyren与舌鳞癌预后的关系及调控机制研究

Long non-coding RNAs (lncRNAs) are known to play key roles in cancer development and progression. In our previous study, we found that miR-195, a member of the miR-15/16 family, has anti-tumor effects in tongue squamous cell carcinoma (TSCC) and significantly correlated with patients mortality. However, the mechanism for miR-195 or miR-15/16 family up-stream regulation remains uncertain. We propose that Cyren, a novel lncRNA, can modulate miR-15/16 family availability by acting as a competitive endogenous RNA (ceRNA). This project is conducive to confirm the prognostic effect of Cyren for patients from TSCC specimens. Trying to investigate the molecular mechanism of Cyren that govern the modulation modes of miR-15/16 family and influence TSCC development and progression by CCK-8, Western blot, dual luciferase reporter assay, etc. Finally, we will verify the “oncogene” effect of Cyren in vivo. This will definitively benefit for understanding the molecular mechanism of TSCC progression and developing biomarkers for TSCC patients based on lncRNA.

长链非编码RNA（lncRNA）在肿瘤进展中起着重要作用。课题组前期研究发现miR-15/16家族成员miR-195在舌鳞癌中发挥抑癌基因的作用，且与患者预后相关，但其上游是否存在调控基因尚不清楚。我们进行的生物学信息分析结果显示，一个新的lncRNA-Cyren转录物上存在miR-15/16家族潜在结合位点，有可能作为竞争性內源RNA（ceRNA）调控miR-15/16家族的表达。为了证实这一假设，本研究拟利用qRT-PCR方法检测舌鳞癌患者组织样本中Cyren表达，分析其与患者预后的相关性；通过CCK-8、Western blot和双荧光报告等实验研究Cyren作为ceRNA调控miR-15/16家族进而影响舌鳞癌发生、发展的分子机制；利用动物体内实验验证Cyren的促癌作用。以上研究将为深入了解舌鳞癌的发病机制和探索判断患者预后的lncRNA分子标志物提供新的思路和实验依据。

长链非编码RNA是新兴的基因调控成员，与癌症的发展有关。本项目用lncRNA表达谱芯片检测了9对肝癌及癌旁组织，发现92个在舌癌组织中异常表达的lncRNA，从中选取差异表达倍数较高的lncRNA-Cyren作进一步研究。利用实时荧光定量PCR检测了120对舌癌及癌旁组织中Cyren的表达，其中lncRNA-Cyren在舌癌中显著下调。Kaplan-Meier 分析发现，Cyren的下调表达与舌癌癌病人的总生存期相关，Cyren高表达的患者预后较差。体外实验结果表明，在舌癌细胞系中过表达Cyren，可以促进细胞增殖。体内实验结果表明，过表达Cyren可以促进裸鼠体内肿瘤形成。机制研究结果表明，Cyren转录本序列中含有潜在的miR-15/16家族（miR-15a/b/195/16/424/497）的结合位点，可通过结合这些miRNA进而调控其靶基因，如Cyclin D1, CDK6, E2F3和Bcl-2的表达。上述研究将为探索预测舌癌患者预后的生物标志物、开发以Cyren为靶点治疗舌癌的有效药物提供新的思路和实验依据，对于提高舌癌的诊断和治疗水平具有重要的理论意义和潜在的临床应用价值。