PTBP3介导的CXCR4可变剪接调控甲状腺乳头状癌侵袭转移的作用及机制研究

伴淋巴结转移的甲状腺乳头状癌（PTC）患者预后欠佳。趋化因子受体CXCR4是调控细胞迁移的重要因子。我们前期发现CXCR4通过可变剪接产生的不同转录本在是否伴淋巴结转移的PTC组织中存在差异表达，体内外实验发现CXCR4可变剪接参与PTC侵袭转移的调控,且可能受剪接因子PTBP3调节。由此，我们提出“PTBP3介导的CXCR4可变剪接调控PTC侵袭转移”的假说。围绕该假说，拟进行以下研究：1、通过免疫共沉淀、RNA pull-down、ceRNA、CHIRP、转录组测序探讨CXCR4可变剪接调控PTC侵袭转移的功能及分子机制；2、通过生物信息学分析、CRISPR-Cas9探讨PTBP3如何调控CXCR4可变剪接；3、结合临床样本探讨其作为PTC预后及潜在治疗靶点的临床价值。本项目旨在从可变剪接这一新视角，为深入理解PTC侵袭转移的机制和筛选转移性PTC治疗靶点奠定基础。

Papillary thyroid cancer (PTC) with lymph nodes metastasis (LNM) was associated with poor prognosis. Chemokine receptor CXCR4 is a key regulatory factor in cell migration. Our previous research found differential expression of alternative splicing variants of CXCR4 in PTC tissue with or without LNM. Functional experiments showed that CXCR4 alternative splicing was involved in the regulation of invasion and metastasis of PTC in vivo and in vitro. Further study found that CXCR4 alternative splicing might be regulated by splicing factor PTBP3. Based on our preliminary data, we hypothesize that PTBP3 mediated CXCR4 alternative splicing modulates the metastasis of PTC. We plan to validate this hypothesis by the following research: 1) To determine the role and underlying mechanisms of CXCR4 alternative splicing regulating PTC metastasis through immunoprecipitation, RNA pull-down, ceRNA, CHIRP, transcriptome sequencing techniques. 2) To clarify how PTBP3 regulates CXCR4 alternative splicing by bioinformatics analysis and CRISPR-Cas9 techniques. 3) To investigate their clinical values as prognostic indicator and potential therapeutic target of PTC with LNM via clinical samples. From a new perspective – alternative splicing, this study will help lay the foundation for PTC mechanistic investigation and find potential targets for intervention.