PRKD3介导的蛋白磷酸化在肿瘤细胞自噬及侵袭迁移中的机制研究

我们前期研究结果提示PRKD3在乳腺癌、肝癌、前列腺癌中高表达，且与肿瘤增殖、侵袭、迁移等恶性表型有关，该蛋白与YAP1蛋白水平存在相关性，改变PRKD3表达可调控自噬，并影响细胞存活，但其机制及调控的下游通路尚未明确。本项目拟采用细胞及动物模型进行研究，观察敲除及过表达PRKD3对肿瘤生物学效应、自噬水平的作用，并在此基础上加入自噬诱导、抑制剂观察调控自噬对PRKD3功能的影响，阐明PRKD3通过自噬调控肿瘤细胞增殖、侵袭及迁移的作用机理。进一步采用磷酸化蛋白质组学、蛋白质组学质谱筛选PRKD3磷酸化调控的下游蛋白，重点关注与肿瘤和自噬相关的Hippo-Yap、mTOR通路等，并对相关差异表达蛋白进行验证，构建以PRKD3为核心的肿瘤恶性表型分子调控网络，阐述并完善PRKD3作用的分子机制，为肿瘤恶性进展以及临床诊疗提供新的理论依据。

Our previous studies suggested that PRKD3 was highly expressed in breast cancer, liver cancer and prostate cancer, and was related to multiple malignant phenotypes in tumor such as proliferation, invasion and migration. Besides, we also found that PRKD3 was correlated with the expression level of YAP1. Altering the expression level of PRKD3 can regulate the activity of autophagy, thus affecting cell survival. However, the exact mechanism and downstream pathways involved have not been clarified. Our study intends to employ cell and animal models to further observe the influence of PRKD3 on tumor biological effect and autophagy after knocking out or overexpression of PRKD3. Moreover, we will also observe the impact of autophagy regulation on PRKD3 function by adding autophagy inducers or inhibitors. Based on these, we will elucidate the regulatory mechanism of PRKD3 in regulating proliferation, invasion and migration in tumor cells through autophagy. Furthermore, the downstream proteins regulated by PRKD3 phosphorylation will be screened by using phosphorylated proteomics and proteomics mass spectrometry, particularly focusing on Hippo-Yap and mTOR pathways which concerned with tumors and autophagy, and the differentially expressed proteins will be verified by animal tissue samples and clinical samples. For the above research, we can construct the molecular regulatory network of tumor malignant phenotype with PRKD3 as the core, and further elaborate the molecular mechanism of PRKD3 which providing the new theoretical basis for the malignant tumor progression and clinical diagnosis and treatment.