PPP3CB抑制Twist1介导的上皮间质转化影响肾癌转移的机制研究

Metastasis is the leading cause of death for renal cell carcinoma (RCC) patients. Therefore, it is particularly important to elucidate the molecular mechanism underlying RCC progression and metastasis in order to improve patient outcomes. It is well known that epithelial-mesenchymal transition (EMT)is a crucial regulator of tumor metastasis. Our previous study showed that PPP3CB could inhibit migration of renal cancer cells via disrupting EMT process and cytoskeleton. Moreover, PPP3CB also interacted with Twist1 which is a critical regulator of EMT. Collectively, these results suggested that PPP3CB mediated tumor metastasis by inhibiting Twist1-induced EMT in RCC. The main content is as follow: the effect of PPP3CB on EMT of RCC cell will be assessed by detecting the RCC cell morphology, cell migration and the expression of EMT markers at cell and animal level; PPP3CB regulating Twist1 and the underlying biological mechanism will be analyzed using mass spectrometry and co-immunoprecipitation assay, meanwhile, immunofluorescence, realtime PCR and western blot will be applied to identify the molecular mechanism by which Twist1 controls the EMT of RCC cell. Thus, our study is expected to illuminate the mechanism of PPP3CB regulating EMT via Twist1 in RCC cell. It will help us find a promising therapeutic target for RCC treatment.

转移是肾癌致死的主要原因之一，深入研究肾癌转移的分子机制对于指导治疗以提高肾癌患者生存率具有重要意义。上皮间质转化（EMT）是肿瘤转移的关键因素之一。我们前期研究发现PPP3CB能抑制肾癌细胞的EMT过程，改变细胞骨架，干扰细胞的迁移；同时PPP3CB能与EMT转录关键因子Twist1相互作用，提示PPP3CB可能通过Twist1调控EMT抑制肾癌的转移。本项目拟在细胞和动物水平检测PPP3CB对肾癌细胞形态、转移能力及EMT标志物的影响，明确PPP3CB对肾癌细胞EMT的调控作用；利用质谱分析和免疫共沉淀确定PPP3CB调控EMT转录关键因子Twist1,同时利用免疫荧光、实时荧光定量PCR和免疫印迹等方法明确PPP3CB调控Twist1影响肾癌细胞EMT的分子机制。本项目旨在阐明PPP3CB调控Twist1介导的EMT在肾癌转移过程中的作用和机制，为抑制肾癌转移发掘新的调控靶点。

癌症是导致人类死亡的最主要原因之一，而上皮间质转化（EMT）和代谢异常均是肿瘤发生发展的关键因素，深入探究其潜在的分子机制对于指导治疗以提高癌症患者生存率具有重要意义。本研究证明PPP3CB能抑制肾癌细胞的EMT过程，改变细胞骨架，干扰细胞的迁移；同时PPP3CB能与EMT转录关键因子Twist1相互作用，并影响其蛋白表达和磷酸化水平，敲低PPP3CB可上调Twist1蛋白水平促进肾癌细胞EMT，揭示PPP3CB可能通过调控Twist1参与肾癌的转移；此外，PPP3CB也能与PDHK1互作并通过影响其活性参与调控膀胱癌细胞的糖代谢过程，综上，我们的研究进一步丰富了钙调磷酸酶PPP3CB在肿瘤中的作用，并探究了肿瘤发生发展新的调控机制，为肿瘤的诊疗提供了新的研究思路和靶点。