YB-1/miR-155通路在人喉癌中的作用及机制研究

YB-1 and miR-155 take part in progresses of many neoplasms, and be expected to be new markers of tumor diagnosis and targets of treatment. So far there is no related research of YB-1 and miR-155 in laryngeal carcinoma. Our previous study found that miR-155 played a promoting role in the development of laryngeal squamous cell carcinoma. Our further research showed significant correlation between YB-1，c-myb and miR-155 in laryngeal squamous cell carcinoma. We research the effect of YB-1/ c-myb/miR-155 pathway in laryngeal carcinoma, enrich the progression mechanism of laryngeal carcinoma, and identify markers of diagnosis, and provide a theoretical basis for exploring new treatment targets. From the basic research aspect, we identify the exist of the pathway through cell transfection, and search the effect of overexpression of YB-1 through experiment in vitro and in vivo, and find downstream target geen through geen chip and dual luciferase assays. The effect and regulator mechanism of YB-1/ c-myb/miR-155 pathway will be identified by the suject. From the clinical research aspect,the relationship between YB-1/c-myb/miR-155 pathway and prognosis will be valued by follow-up, and this will provide new theory evidence for searching new treatment target of laryngeal carcinoma.

YB-1和miR-155参与了多种肿瘤的发生，并有望成为新的肿瘤诊断标记物和治疗靶点。目前尚无喉癌中YB-1和miR-155的相关研究。本项目的前期研究已经证实miR-155在喉癌的发生发展中起到关键的促进作用,而 YB-1，c-myb,miR-155三者在喉癌中的表达具有显著的相关性。我们拟以YB-1/ c-myb/miR-155通路在喉癌的作用为切入点，丰富喉癌的发生演进机制、认证新的诊断标记物，并为发掘喉癌新的治疗靶点提供理论依据。项目从基础研究角度通过细胞转染证实该通路在喉癌中的存在，通过离体和在体实验证实YB-1在喉癌发生发展中的作用，通过基因芯片、双荧光素酶实验寻找该通路下游靶基因，全面阐述YB-1/ c-myb/miR-155通路的调控机制及功能。从临床角度，通过随访评估YB-1/c-myb/miR-155通路与喉癌患者预后的关系，为寻找喉癌治疗靶点提供新的理论依据。

喉癌是常见的头颈部恶性肿瘤，是东北地区最常见的头颈部恶性肿瘤。本课题通过对120例喉癌标本，40例粘膜标本进行western blot和real-time PCR检测。分别建立YB-1稳定高表达和低表达hep-2细胞系，通过CHIP确定YB-1，c-myb和miR-155之间的调控关系。通过transwell小室MTT法考察YB-1/miR-155通路对体外的喉癌细胞生长、转移和侵袭能力的影响。通过生物信息学方法寻找到miR-155靶点，应用western blot确定，探讨YB-1及miR-155与临床病理参数的以及喉癌生存期的关系。我们证实了YB-1/miR-155通路在喉癌组织中表达上调，确定了YB-1，c-myb和miR-155之间的调控关系，明确了该通路的调控机制。明确了YB-1/miR-155通路在体外对喉癌细胞的侵袭和转移的促进作用。找到并验证了miR-155的新靶点TGFβ1，进一步诠释了该通路在喉癌的作用机制。分析YB-1及miR-155表达量和临床病理参数的关系，发现了YB-1，miR-155在T3T4期喉癌和中低分化喉癌的表达显著高于T2期喉癌和高分化喉癌。YB-1蛋白、miR-155表达和患者生存期显著正相关。本研究首次证实了YB-1/miR-155通路的存在，明确了该通路在喉癌在喉癌发生发展中的促进作用及机制，发现了YB-1蛋白和不良预后的相关性。本研究为进一步明确喉癌发生发展机制，提供新的预后指标提供了证据。