非编码RNA：LncRNA和miRNA在喉癌中的作用及其对表观遗传和信号传导的调控

Laryngeal cancer is a malignant tumor which is seriously threatening to human health. Its mortality is still high,especially for later stage of laryngeal cancer. The precise molecular mechanisms of laryngeal cancer have not been elucidated.It is mainly considered to related with oncogene and tumor suppressor gene disorders. Non coding RNAs are RNAs that not translated to protein. Our early study shows non-coding-LncRNA and miRNA are abberantly expression in laryngeal cancer. But their regulation in epigenetic and signal transduction is unclear. Our study further suggested LncRNA-HOTAIR can modulate PTEN gene promoter methylation and miR-21 can regulate the expression of Ras. Thus we hypothesized that LncRNA and miRNA should regulate the occurrence and development of laryngeal carcinoma through epigenetic and signal transduction mechanism. To confirm this hypothesis, we will use the methods of microarray, qRT-PCR, apoptosis assay, Western-blot, Chip and RNA-chip， RNA-pull-down and bioinformatics analysis methods to observe the role of LncRNA and miRNA in laryngeal cancer and clarify the mechanism of laryngeal cancer development from a new perspective. This study is expected to provide a new effective target and strategy for therapy of laryngeal cancer.

喉癌是严重威胁人类健康的疾病，尤其是晚期喉癌死亡率仍较高。喉癌发生发展的确切分子机制尚未阐明，多认为与癌基因和抑癌基因的失调有关。非编码RNA指的是不被翻译成蛋白质RNA。本课题组的前期研究筛选出喉癌中差异表达的非编码LncRNA和miRNA，但它们在喉癌中作用，以及对喉癌中表观遗传和信号传导的调控尚不清楚。课题组的预实验发现LncRNA-HOTAIR能调节PTEN基因的启动子甲基化，miR-21能调节Ras的表达。由此我们推测LncRNA和miRNA将可能通过调节表观遗传、信号传导或与蛋白结合等机制调控喉癌的发生发展。为证实这一假说，我们将运用microarray，定量PCR，凋亡实验，蛋白印迹，Chip，RNA免疫共沉淀，pull-down和生物信息分析等方法，观察LncRNA和miRNA在喉癌中的作用，并从新的视角阐明喉癌发生发展的机制。本研究将可望为喉癌治疗提供新的有效靶点和策略。

喉癌是严重威胁人类健康的疾病，尤其是晚期喉癌死亡率仍较高。喉癌发生发展的确切分子机制尚未阐明，多认为与癌基因和抑癌基因的失调有关。非编码RNA指的是不被翻译成蛋白质RNA。本课题组的前期研究筛选出喉癌中差异表达的非编码LncRNA和miRNA，但它们在喉癌中作用，以及对喉癌中表观遗传和信号传导的调控尚不清楚。本课题探讨非编码RNA：miRNA和lncRNA在喉癌中的作用及其机制，其主要作用是寻找喉癌的分子标志和可能的治疗靶点。通过microarray，定量PCR等方法，我们筛选出了喉癌中起到关键分子marker作用的miRNA是：miR-19a，miR-205等。喉癌中起到重要分子marker作用的LncRNA是：HOTAIR，NEAT1，H19，RP11和Let。进一步的机制研究发现这些miRNAs和LncRNA均在喉癌中扮演癌基因或抑癌基因的角色。miR-19a能促进喉癌肿瘤细胞的生长，与其调节TIMP-2有关。miR-205能抑制肿瘤的生长并促进其凋亡，与调控Bcl2有关。HOTAIR能促进喉癌细胞的生长，与调控TPEN甲基化有关。NEAT1通过调节miR-107来调控CDK6，从而促进喉癌细胞的增殖。H19通过调控miR-148a来调控DNMT1，从而调控喉癌的整体甲基化水平。通过本课题的研究，进一步丰富了喉癌的发生发展的机制，为喉癌的治疗靶点的筛选，提供了理论和实验依据。