HOXB9靶向MMP12促进喉鳞状细胞癌增殖及侵袭的机制研究

喉鳞状细胞癌是头颈部最常见的恶性肿瘤之一，尽管在过去20年中治疗已不断改善，但死亡率仍然很高。关于喉癌细胞的增殖及侵袭机制，目前仍未完全阐明。申请者在前期工作中，通过基因芯片筛选出部分HOX基因家族成员在喉癌组织中表达增高，进一步PCR及大样本组织芯片验证，发现HOXB9与喉癌患者临床病理分级及预后高度相关。后续实验通过CRISPR/CAS9技术敲除HOXB9后发现，喉癌细胞增殖及侵袭能力、体内裸鼠成瘤能力均显著降低。通过敲除HOXB9后下游基因芯片筛选，结合生物信息学工具分析，推测HOXB9可能通过靶向结合MMP12,促进喉癌细胞的增殖及侵袭。本项目拟在此基础上，从大样本组织芯片中分析HOXB9和MMP12的共表达和定位，以及二者共表达对喉癌患者预后的判断价值。同时，分别在细胞和动物水平进一步探讨二者的调控关系及调控模式和位点，为寻找有效的抑制喉癌细胞增殖和侵袭靶点提供理论依据。

Laryngeal squamous cell carcinoma (LSCC) is one of the most common malignant tumors of the head and neck. Although treatment has been improving over the past 20 years, the mortality rate is still very high. The mechanism of proliferation and invasion of laryngeal cancer cells has not been fully elucidated. In the previous work, some members of HOX gene family were screened by gene chip to increase their expression in laryngeal cancer tissues. Further verification by PCR and large sample tissue chip showed that HOXB9 was highly correlated with clinical pathological grading and prognosis of laryngeal cancer patients. In the follow-up experiments, HOXB9 was knocked out by CRISPR/CAS9 system. It was found that the proliferation and invasive ability of laryngeal cancer cells and tumorigenicity of nude mice in vivo were significantly reduced. By screening the downstream gene chips after HOXB9 knockout and analyzing with bioinformatics tools, it is speculated that HOXB9 may promote the proliferation and invasion of laryngeal cancer cells by targeting MMP12. On this basis, we will analyze the co-expression and localization of HOXB9 and MMP12 in large sample tissue microarray, and the value of co-expression of HOXB9 and MMP12 in predicting the prognosis of laryngeal cancer patients. At the same time, we further explored the regulatory relationship, regulation pattern and locus of the two molecule at cellular and animal levels, which could provided a theoretical basis for finding effective targets for inhibiting proliferation and invasion of laryngeal carcinoma cells.