CRMP4转录调控与前列腺癌转移机理的研究

Ninety percent of cancer-related mortality is caused by metastases. The expression loss of metastasis suppressor gene is closely related to cancer metastasis. The applicant has confirmed for the first time that CRMP4 is a prostate cancer metastasis suppressor gene and the promoter DNA methylation of CRMP4 is implicated to its expression loss ( Oncogene, 2010 ). The study was supported by the 2009 National Youth Fund of Chinese National Nature Science Foundation. This achievement has been awarded the 2011 Guangdong Province Science and Technology Progress Award # 1 and the 2012 Chinese National Science and Technology Progress Award # 1. Based on the original research, this project will continuously be carried out as follows. The binding transcription factors of CRMP4 and its transcriptional regulation mechanisms will be explored using bioinformatics methodology analysis and other experiments such as CHIP, EMSA and reporter gene assay. Espeacially, how the transcription factors c-Myc affects on CRMP4 transcriptional activity before and after the DNA methylation teatment of CRMP4 promoter will be identified. Subsequently, the protein interacted with CRMP4 and the signal pathway controlled by CRMP4 will be confirmed using the molecular biological techniques including tandem affinity purification, mass spectrometric analysis, immunoprecipitation and immunoblotting technique.Finally, the analysis and authentication of CRMP4 functional domains will be carried out and the key proteins of CRMP4 binding will further be identified. The study will clarify the mechanisms of CRMP4 transcriptional regulation and the key molecular events of the CRMP4 inhibition of prostate cancer metastasis, which will provide an effective diagnosis and intervention target for early metastasis of prostate cancer.

肿瘤转移是90%的癌症相关死亡的原因，转移抑制基因表达沉默与肿瘤转移密切相关。申请者获2009年国家自然科学青年基金资助研究证实CRMP4为前列腺癌转移抑制基因，启动子甲基化为其表达沉默的调控机制（Oncogene, 2010），该成果先后获得2011年广东省科技进步一等奖和2012年国家科学技术进步一等奖。本项目在原创性研究的基础上开展延续性研究，采用生物信息学方法、CHIP、EMSA和报告基因质粒等实验探索与CRMP4启动子结合的转录因子和转录调控机制，特别是c-Myc对CRMP4启动子甲基化前后转录活性的调控；利用串联亲和纯化技术、质谱分析、免疫共沉淀和免疫印迹等技术探明CRMP4相互作用的蛋白及其调控的信号通道；分析并认证CRMP4功能结构域及与之结合的关键蛋白。阐明CRMP4转录调控及其抑制前列腺癌转移过程的关键分子事件，为干预前列腺癌早期转移提供有效的靶标和理论依据。

肿瘤转移是90%的癌症相关死亡的原因，转移抑制基因表达沉默与肿瘤转移密切相关。申请者获2009年国家自然科学青年基金资助研究证实CRMP4为前列腺癌转移抑制基因，启动子甲基化为其表达沉默的调控机制（Oncogene, 2010），该成果先后获得2011年广东省科技进步一等奖和2012年国家科学技术进步一等奖。随后采用生物信息学方法、CHIP、EMSA和报告基因质粒等实验探索与CRMP4启动子结合的转录因子和转录调控机制，特别是c-Myc对CRMP4启动子甲基化前后转录活性的调控；利用串联亲和纯化技术、质谱分析、免疫共沉淀和免疫印迹等技术探明CRMP4相互作用的蛋白及其调控的信号通道；分析并认证CRMP4功能结构域及与之结合的关键蛋白。阐明了CRMP4转录调控及其抑制前列腺癌转移过程的关键分子事件，为干预前列腺癌早期转移提供有效的靶标和理论依据。该课题的研究发表于《Oncotarget》、《Prostate》、《Int J Clin Exp Pathol》等上，该课题系列研究发表相关研究SCI论文4篇，获得国家发明专利1项。