磷酸甘油酸激酶1（PGK1）通过Src及下游通路调控口腔鳞癌侵袭转移的机制研究

Invasion and metastasis are the primary factors in the failure of oral squamous cell carcinomas(OSCCs) therapy, and the current understanding of it has yet to be further in-depth. Glycolytic metabolism kinases are closely associated with tumor invasion and metastasis, while it is less known in OSCCs. After screening the expression of multiple metabolic kinases in OSCCs cells with different abilities of invasion and migration, we found that the exprerssion of phosphoglycerate kinase 1 (PGK1) was closely related to invasion and migration of OSCCs cells. And then we clarified the role of PGK1 through cell function test. To further clarify the mechanism of PGK1in invasion and migration of OSCCs cells, we preferred protein kinases phosphorylation chip screening, by which it was found that Src and its downstream signal pathway was closely associated with the expression level of PGK1. This result was verified by Western-blot. As we hypothesized that PGK1 might regulate the invasion and metastasis of oral squamous cell carcinomas through Src and its signaling pathways. The topics will be expanded on the level of molecular, cellular, animal experiment in order to explore the role of Src and its downstream pathway in the regulation of PGK1 on invasion and metastasis of OSCCs cells. Then we study the correlation between the expression of PGK1 and metastasis and prognosis in clinical OSCCs tissues of patients. Based on the above researches, we hope to clarify the mechanism of action and regulation of PGK1 in invasion and metastasis of OSCCs and provide the theoretical basis for PGK1 as a molecular marker of metastasis and prognosis of OSCCs.

侵袭转移是口腔鳞癌治疗失败的首要因素，其分子调控机制尚未完全阐明。糖酵解代谢激酶与肿瘤侵袭转移密切相关，在口腔鳞癌中研究尚不深入。为研究代谢酶在口腔鳞癌中的作用，我们检测了不同迁移能力口腔鳞癌细胞中激酶表达水平，发现磷酸甘油酸激酶1（PGK1）和肿瘤细胞侵袭、迁移密切相关，并通过细胞功能试验明确了其作用。为阐明PGK1作用机制，我们采用蛋白磷酸化芯片筛查、Western-blot验证等方法，初步明确转移关键分子Src及其下游通路活性受PGK1所调控。据此推测PGK1可能通过Src通路调控口腔鳞癌侵袭转移。课题将以此展开，在分子、细胞、动物水平实验进深入研究Src及下游通路在PGK1对口腔鳞癌侵袭转移调控中的作用，并在临床标本组织中研究PGK1和口腔鳞癌转移和预后的相关性，旨在阐明PGK1调控口腔鳞癌侵袭转移的作用及机制，为PGK1作为口腔鳞癌转移及预后判断的分子标志物提供理论依据。

侵袭转移是影响口腔鳞癌预后的一个重要因素，但其分子调控机制尚未完全阐明。在前期研究和文献调研的基础上，本研究先以糖酵解代谢激酶磷酸甘油酸激酶1（PGK1）为切入点，进一步研究发现另一种细胞有氧糖酵解代谢酶——磷酸甘油酸酸变位酶1（PGAM1）也具有调控口腔鳞癌细胞侵袭、迁移的作用。进一步利用蛋白磷酸化芯片和Western-blot等方法，并明确PGAM1可能通过Paxillin/FAK/SRC信号转到通路来影响肿瘤细胞运动迁移转。随后，在临床组织样本中发现PGAM1的表达水平和口腔鳞癌淋巴结转移有关，这些研究为PGAM1作为口腔鳞癌转移及预后判断的分子标志物提供理论依据。此外，本课题组成员还充分利用基金支持参与了利用肿瘤原代细胞模型开展化疗药物和靶向药物敏感性的研究，以及第二代基因测序技术在体细胞基因变异临床检测中的应用研究。这些研究为进一步推进肿瘤原代模型和第二代基因测序技术在转化研究和临床诊断中的应用奠定了基础。