LncRNA与miRNA在喉癌中的相互靶向调控作用及其机制

Laryngeal cancer is a serious threat to human health. In advanced laryngeal cancer, mortality rate is high and lack effective treatments. The development of laryngeal carcinoma is related to disor-ders in oncogenes and tumor suppressor genes. Our previous studies involved screening differentially expressed long non-coding lncRNA and miRNA in larynx tissue. The results indicated the involvement of lncRNA and miRNA in the initial progression of laryngeal cancer. However, it is unclear how lncRNA and miRNA expression are regulated or co-regulated in the development of laryngeal carcinoma. Our research group found that reduced expression of miR-21 can regulate the gene ras, and the expression of MMP-2 which promotes cell migration. Meanwhile, lncRNA-HOTAIR regulates the PTEN gene promoter via methylation. Other tumors with the presence of both lncRNA and miRNA showed mechanisms of co-regulation. Therefore, we hypothesize lncRNA and miRNA are co-regulated in laryngeal carcinoma by possible epigenetics and signal transduction mechanisms. To confirm this hypothesis, we will use the method of gene chips, quantitative PCR, methylation detection, bioinformatics analysis, observation lncRNA and mutual targeting miRNA. The results from this study is expected to contribute to the development of a novel and effective treatment for the laryngeal carcinoma.

喉癌是严重威胁人类健康的疾病，中晚期喉癌死亡率仍较高，缺乏有效的治疗手段。喉癌发生发展与癌基因和抑癌基因的失调相关。本课题组前期研究筛选出喉癌中差异表达的非编码LncRNA和miRNA，并初步阐明但它们在喉癌进展中的作用，但LncRNA和miRNA之间是否在喉癌中存在相互靶向调控作用及其机制尚不清楚。课题组发现下调miR-21能调节Ras的表达，并通过调节MMP-2 表达促进细胞迁移。同时，LncRNA-HOTAIR能调节PTEN基因的启动子甲基化。而在其他肿瘤中发现了LncRNA和miRNA存在相互靶向调控作用，由此我们推测LncRNA和miRNA将可能通过相互靶向调节表观遗传、信号传导等机制调控喉癌的发生发展。为证实这一假说，我们将运用基因芯片，定量PCR，甲基化检测和生物信息分析等方法，观察LncRNA和miRNA在喉癌中相互靶向调控的作用，并有望为喉癌治疗提供新的策略。

喉鳞状细胞癌是一种具有高度侵袭性的恶性肿瘤。长链非编码RNA（lncRNA）参与多种疾病的进展，但对喉鳞状细胞癌进展的调控机制尚不清楚。在这项研究中，我们报告了lncRNA H19在喉癌中显著过表达, lncRNA H19的表达水平与喉鳞状细胞癌患者的生存率呈负相关。本课题在分析分析其病理特征的基础上，构建lncRNA H19慢病毒表达载体，发现沉默lncRNA H19表达抑制喉鳞状细胞癌细胞迁移，侵袭和增殖。在体内裸鼠移植瘤实验中，也证实沉默lncRNA H19抑制裸鼠移植瘤的生长。我们鉴定了miR-148a-3p作为H19的抑制靶基因。miR-148a-3p的过度表达减少了喉鳞状细胞癌细胞迁移，侵袭和增殖，而抑制miR-148a-3p的表达则相反。由此可见，lncRNA H19沉默抑制喉鳞状细胞癌进展需要miR-148a-3p的活性。最后，我们还鉴定了DNA甲基转移酶DNMT1作为miR-148a-3p的靶基因，发现了miR-148a-3p过表达和lncRNA H19沉默都抑制了细胞DNA甲基化水平。总之，我们的研究表明lncRNA H19通过miR-148a-3p和DNMT1促进了喉鳞状细胞癌进展。为喉肿瘤发病机制的研究和新靶点的发现提供了新的思路以及一定的理论依据。