CPEB4介导的翻译调控在鼻咽癌转移过程中的作用及机制研究

The aberrant transcripts of genomes were considered to be the primary cause during the development of malignancies. Post-transcription process has being taken increasingly seriously, the regulation of post-translational modifications mediated by protein was widely studied.In eukaryotic cells, there were a large number of dormant mRNA containing CPE sequence, and these mRNAs could be activated by the CPE binding protein and started the process of transcription, and during the process of regulation of mitosis and neurodevelopmental, this process of dormant mRNA containing CPE sequence being activated played a crucial role.Many researches suggested translational regulation mediated by CPE binding protein 4(CPEB4) was closely related to the proliferation, angiogenesis and invasion of tumor, our previous studies indicated that the expression of CPEB4 increased obviously in nasopharyngeal carcinoma (NPC) tissue. New research method iTRAQ and normal methods such as gene transfection and animal model were chosen for the research of CPEB4 regulation of the lymph nodes metastasis of NPC, the key signaling pathways or molecules in the downstream of CPEB4 were expected to be found, and it is also hoped that the findings could offer a new perspective and possibility for the prevention and treatment of the lymph nodes metastasis of NPC.

目前，蛋白质介导的翻译调控在真核细胞各种生物学过程中发挥的作用逐渐受到研究者的关注。真核细胞中有着数量众多的含有细胞质腺苷化元件（CPE）序列的休眠mRNA，它可以在CPE结合蛋白（CPEB）蛋白作用下激活其翻译过程，该过程在真核细胞的有丝分裂、神经细胞生长及早期胚胎发育过程中发挥十分关键的作用。研究发现CPEB蛋白家族成员CPEB4介导的翻译调控与部分肿瘤细胞的增殖、血管形成及侵袭密切相关，但仅限于mRNA水平的研究，且其中的具体分子机制尚不明确，我们前期的研究发现CPEB4蛋白在鼻咽癌组织中表达显著增高，且与鼻咽癌患者的N分期有显著的相关性。本课题拟采用基因转染、同位素标记的相对与绝对定量等实验技术研究CPEB4在鼻咽癌淋巴结转移过程中的作用，并从蛋白水平明确CPEB4下游的关键信号通路或分子，为鼻咽癌转移的防治研究提供一个新的思路和切入点。

真核细胞中有着数量众多的含有细胞质腺苷化元件（CPE）序列的休眠mRNA，它可以在CPE 结合蛋白（CPEB）蛋白作用下激活其翻译过程，该过程在真核细胞的有丝分裂、神经细胞生长及早期胚胎发育过程中发挥十分关键的作用。研究发现CPEB 蛋白家族成员CPEB4 介导的翻译调控与部分肿瘤细胞的增殖、血管形成及侵袭密切相关，但仅限于mRNA水平的研究，且其中的具体分子机制尚不明确，我们前期的研究发现CPEB4 蛋白在鼻咽癌组织中表达显著增高，且与鼻咽癌患者的N 分期有显著的相关性。本研究中我们采用了GEO大数据相关生物信息学统计、基因工程学方法、细胞行为学检测、蛋白印迹法、同位素标记的相对与绝对定量、共聚焦免疫荧光、裸鼠动物模型等实验技术研究CPEB4在鼻咽癌发生发展及转移过程中的作用，本研究结果提示CPEB4在鼻咽癌中发挥促癌作用，其高表达与鼻咽癌低分化、转移发生呈正相关。并从蛋白水平明确CPEB4下游的关键信号通路或分子，iTRAQ的结果也提示CPEB4过表达/干扰后调控的下游分子大多聚集在参与鼻咽癌分化、转移等生物学过程，此外，裸鼠移植瘤实验对CPEB4过表达后的鼻咽癌细胞成瘤效果进行了评估，过表达CPEB4后移植瘤体积显著增大。经过筛选本研究进一步分析了CPEB4下游分子CTNNB1的表达，通过一系列分子生物学手段证实两者的相互作用，并拟在进一步的研究中将两者的作用在裸鼠动物模型中进行验证，本研究的成果为鼻咽癌的防治研究提供一个新的可靠的切入点。