侵袭力是肿瘤细胞实现转移的关键。我们筛选出中国人前列腺癌差异表达miRNA谱（BMC Genomics，2013），发现并证明miR-224与靶基因TRIB1对侵袭力起关键调控作用（Int J Cancer，2014），进一步发现TRIB1与肿瘤上皮间质化（EMT）相关，但miR-224/TRIB1调控肿瘤EMT的下游靶点和机制是什么？通过高通量筛选到一批与TRIB1相关的Wnt和MAPK通路基因，为此提出“TRIB1结合Wnt/MAPK通路的关键蛋白，激活Wnt/MAPK，诱导肿瘤EMT调控前列腺癌侵袭力”的假设。拟用CRISPR/Cas9基因敲除等技术，获得miR-224/TRIB1调控EMT的下游靶点，通过免疫共沉淀，验证TRIB1与靶点的结合激活机制，最终阐释TRIB1激活MAPK/Wnt诱导肿瘤EMT调控侵袭力的分子机制，为前列腺癌侵袭力的诊断和治疗新靶点提供理论依据。

Tumor invasion is a key factor for the metastasis of patients with prostate cancer. For the first time, we found differential expression profile in PCa samples of Chinese patients by miRNA microarray analyses (BMC Genomics,2013).We demonstrated that miR-224 acts as a tumor suppressor by regulating its target oncogene TRIB1, and miR-224-TRIB1 axis represents a key regulatory mechanism to PCa invasion(Int J Cancer,2014). But what is the target and regulatory mechanism that miR-224/TRIB1 regulate EMT of tumor? Based on the Wnt and MAPK pathway gene combinations which is connected with TRIB1 acquired by high-throughput screening,we hypothesized that miR-224-TRIB1 axis induces EMT by interacting with key genes and activating MAPK & Wnt pathways, plays a crucial role in regulating prostate tumor invasion.We get the downstream targets that miR-224/TRIB1 regulat EMT by the gene knock-out technology of CRISPR/Cas9.CoIP experiment is intent to be carried out to verify the mechanism and effect among miR-224 and its targets,MAPK/Wnt signaling pathways and EMT on PCa invasion, and further figure out the targeting genes or pathways involving the tumor EMT and invasion of miR-224-TRIB1 axis, and provide a scientific basis for anti-tumor invasion judgement and therapy of prostate cancer.

我们前期研究中筛选出中国人前列腺癌差异表达miRNA谱，发现并证明miR-224与靶基因TRIB1对侵袭力起关键调控作用，进一步发现TRIB1与肿瘤上皮间质化相关，但miR-224/TRIB1调控肿瘤EMT的下游靶点和机制是什么？通过高通量筛选到一批与TRIB1相关的Wnt和MAPK通路基因，为此提出“TRIB1结合Wnt/MAPK通路的关键蛋白，激活Wnt/MAPK，诱导肿瘤EMT调控前列腺癌侵袭力”的假设。项目总体上达到研究目标，我们通过构建过表达细胞株以及细胞功能实验的研究，验证了TRIB1调节前列腺癌细胞生长和转移的现象，利用过表达TRIB1裸鼠的肿瘤种植实验，明确了TRIB1促进前列腺癌的作用。同时通过高通量筛选，明确TRIB1促进前列腺腺体细胞分泌趋化因子调节免疫系统对肿瘤的影响。在研究过程中，我们还发现另外2个基因HOXD10和ASPM对前列腺癌的进展有促进作用。HOXD10在前列腺增生样本中高表达，在前列腺癌组织中的表达明显低于前列腺增生组织，且随着 Gleason Score 越低，阳性表达越弱。此外，抑制ASPM的表达有助于减缓前列腺癌的进展并且有利于患者的预后。以上结果为进一步研究肿瘤免疫抑制作用提供基础，继续完善相关下游基因的研究，构建相关基因调控网络。