CDK15和CCNY双向调控促进结直肠癌发生发展的机制研究

结直肠癌是我国常见的消化道恶性肿瘤，由于缺乏有效的治疗靶点，其治疗效果并不理想。我们前期的研究结果发现，CDK15在人结直肠癌组织和细胞中均高表达，并且与患者的生存期呈负相关；降低CDK15能够有效抑制结直肠癌的增殖并将细胞周期阻滞在G2/M期；体内裸鼠实验和PDX模型得到了相似的结果；Pull down实验显示CCNY可能是CDK15潜在的靶分子；体外激酶实验发现CDK15可以磷酸化CCNY，并且CCNY又可以促进CDK15的激酶活性，提示结直肠癌中CDK15和CCNY可能通过相互调节形成正反馈，从而促进结直肠癌的生长。本项目拟通过iTRAQ定量蛋白组学，免疫共沉淀，shRNA技术和基因敲除小鼠等，进一步研究CDK15-CCNY-X轴在结直肠癌发生发展中的作用；同时将利用超级计算机筛选出针对CDK15的小分子抑制剂，体内外验证其疗效，为以CDK15为靶点的结直肠癌防治提供新的实验依据。

Colorectal cancer is a common gastrointestinal cancer in China. Due to the lack of effective treatment targets, its treatment effect is not ideal. Our previous study found that CDK15 was highly expressed in human colorectal cancer tissues and cells, and negatively correlated with the survival time of patients; reducing CDK15 could effectively inhibit the cell proliferation of colorectal cancer and block the cell cycle in G2 / M phase; similar results were obtained in in vivo xenograft mice model and PDX model; pull down assay showed that CCNY may be a potential molecule target of CDK15. Our study further found that CDK15 could phosphorylate CCNY in vitro, and CCNY could promote the kinase activity of CDK15, suggesting that CDK15 and CCNY could form positive feedback through mutual regulation, so as to promote colorectal cancer growth. This project intends to further study the role of CDK15-CCNY-X axis in the occurrence and development of colorectal cancer through iTRAQ quantitative proteomics, immunocoprecipitation, shRNA technology and gene knockout mice, etc.; at the same time, we will use supercomputer to screen out small molecular inhibitors for CDK15, verify their efficacy in vitro and in vivo, and provide new experimental basis for the prevention and treatment of colorectal cancer in the clinic.