MAP3K10/DYRK2介导Hedgehog旁路途径活化对胰腺癌干细胞生物学行为影响的研究

Pancreatic cancer stem cells(PCSCs) are malignant core cells of pancreatic cancer and contribute to the initiation, progression and prognosis of pancreatic cancer.Hedgehog (Hh) signaling is one of the most crucial pathways of PCSCs that aberrantly activated.Although the classic blocker of Hh signaling could reduce the proportion of PCSCs in pancreatic cancer,it could not kill the PCSCs completely.Through our previous research we find that MAP3K10/DYRK2 are activated abnormally in PCSCs,which may activate the downstream of Hh signaling bypass, by which PCSCs escape chemotherapy.The objective of our research is to confirm the expression levels of MAP3K10/DYRK2 in pancreatic cancer cells and PCSCs and its relationship with clinicopathologic degree,to verify the activation of Hh signaling by MAP3K10/DYRK2 through IP and BiFC,to detect the effects of MAP3K10/DYRK2 on the malignant biological behaviors of pancreatic cancer and the maintenance of PCSCs, to exploit new blockers targeting MAP3K10/DYRK2 proteins and confirm their pharmacological effect on PCSCs.Our project may gain a clear idea of the molecular mechanisms of PCSCs escaping chemotherapy and may be crucial for the development of new targetd agents.It may lay a foundation for the improvement on treatment effect of pancreatic cancer.

胰腺癌干细胞是胰腺癌恶性核心细胞，决定了胰腺癌的发生、发展和转归。Hedgehog信号通路是胰腺癌干细胞中最重要的异常激活通路，其阻滞剂能有效的降低胰腺癌干细胞比例，但并不能彻底的杀灭胰腺癌干细胞。我们前期研究发现MAP3K10/DYRK2在胰腺癌干细胞中异常激活，可能作为Hedgehog旁路途径激活下游信号通路而导致胰腺癌干细胞逃逸治疗。本研究拟在前期研究基础上：明确MAP3K10/DYRK2在胰腺癌及胰腺癌干细胞中的表达及临床病理联系；运用IP和BiFC技术证实MAP3K10/DYRK2对Hedgehog信号通路的激活；检测MAP3K10/DYRK2对胰腺癌恶性生物学行为和胰腺癌干细胞"干性"维持的影响；开发MAP3K10/DYRK2蛋白阻滞性药物，检测其对胰腺癌干细胞的杀灭作用。本项目有望进一步揭示胰腺癌干细胞逃逸当前治疗的分子机制，为提高胰腺癌治疗效果设计合理靶向药物奠定实验基础。

肿瘤干细胞是近年来研究的热点，大量研究已经证实肿瘤干细胞是肿瘤恶性生物学特征的载体。通过本次研究，我们研究组阐明MAP3K10/DYRK2对胰腺癌恶性生物学特征及胰腺癌干细胞“干性”维持的影响。并且进一步阐明Smo/MAP3K10/DYRK2/Gli2四者间靶向关系及对Hh信号通路的调控机制。同时我们也做了部分开发MAP3K10/DRYK2阻滞剂的工作，为胰腺癌临床药物开发打下基础。我们还解决了MAP3K10/DYRK2异常表达是否是胰腺癌干细胞逃逸Hh信号通路阻滞剂治疗的关键原因这一问题。我们前期研究证实MAP3K10/DYRK2在胰腺癌及胰腺癌干细胞中呈异常表达，为进一步实验打下了基础。另外，胰腺癌中Smo/MAP3K10/DYRK2/Gli2四者之间的靶向作用关系。我们前期通过免疫共沉淀技术发现DYRK2靶向作用于Gli2，但四者间的靶向调控关系及作用的具体位点尚未完全阐明清楚，我们通过IP、QIP、BiFC及LC-MS试验充分揭示四者间的靶向作用关系。该项目将胰腺癌干细胞“干性”维持关键MAP3K10/DYRK2作为研究突破点，深入探讨其靶基因的调控。有望阐明胰腺癌干细胞逃逸临床治疗的分子机制，并为彻底杀灭胰腺癌干细胞提供有效分子靶点。开发相关分子的阻滞剂将为临床药物开发打下基础。