TOPK调节Raf-1/MEK/ERK对胰腺神经内分泌肿瘤肝转移的影响及治疗靶点的初步研究

The key point and difficulty of the treatment for pancreatic neoroendocrine neoplasm is the liver metastasis. The occurence of the liver metastasis of pancreatic neoroendocrine tumor is closely related to the open or closure of Raf-1/MEK/ERK pathway of tumor cell, which is proved by our previous studies. The progress of research in TOPK (TLAK cell-originated protein kinase) suggests that TOPK may involve the accommodation of the liver metastasis of pancreatic neoroendocrine neoplasm via the interaction with the Raf-1/MEK/ERK pathway. It means TOPK might be a therapuetic target for the liver metastasis of pancreatic neoroendocrine neoplasm.This project is focused on the interaction of TOPK and Raf-1/MEK/ERK pathway. Using bioinformatic analysis for the phosphorylation site of TOPK, we try to screen and identify the interaction site of TOPK and Raf-1/MEK/ERK pathway, as well as the effect of TOPK to the activity of kinase, to the Raf-1/MEK/ERK pathway, and to the liver metastasis of tumor. We expected to explore the relationship between TOPK, Raf-1/MEK/ERK pathway and liver matastasis of pancreatic neoroendocrine neoplasm on the molecular mechanism level, and to seek the key therapuetic targets for pancreatic neoroendocrine neoplasm.

肝转移是胰腺神经内分泌肿瘤治疗的难点与关键。我们的前期研究证实，胰腺神经内分泌肿瘤是否发生肝转移与肿瘤细胞Raf-1/MEK/ERK通路的开放或关闭密切相关。进一步研究发现，TOPK可能通过与Raf-1/MEK/ERK通路的相互作用参与了胰腺神经内分泌肿瘤肝转移的调节，与Raf-1和ERK位点均存在调节作用，提示TOPK可能是预防和治疗胰腺神经内分泌肿瘤肝转移的治疗靶点。本项目围绕TOPK与Raf-1/MEK/ERK的相互作用展开研究。拟通过对TOPK磷酸化位点的生物信息学分析，筛查、确定TOPK与Raf-1/MEK/ERK相互作用的位点、对激酶活性的影响、对Raf-1/MEK/ERK通路的影响，以及对肿瘤肝转移的影响。我们期望，从分子机制上进一步揭示TOPK、Raf-1/MEK/ERK与胰腺神经内分泌肿瘤肝转移之间的关系，寻找胰腺神经内分泌肿瘤肝转移关键治疗靶点。

胰腺神经内分泌肿瘤是逐年高发的胰腺肿瘤，很多病人在确诊时已经发生了转移，肝转移是最常见的转移部位，研究胰腺神经内分泌肿瘤和肝转移有重要的临床意义和现实价值。TOPK在多种肿瘤中高表达，对肿瘤细胞的增殖、分化、转移等起到调节作用。本研究在胰腺神经内分泌肿瘤BON-1细胞中敲降TOPK，完成了体外表型实验，并完成了转录组测序、Label-free非标记定量蛋白分析、能量代谢产物分析。得出以下结论1.敲降TOPK抑制胰腺神经内分泌肿瘤BON-1细胞系的增殖、克隆形成和侵袭迁移。2.TOPK通过调控MEK/ERK信号通路抑制胰腺神经内分泌肿瘤BON-1细胞系的恶性表型，敲降TOPK抑制BON-1细胞系能量代谢。3.TOPK靶向抑制剂HI-TOPK-032显著抑制BON-1胞系的恶性表型4.在胰腺神经内分泌肿瘤组织中,TOPK阳性表达与肿瘤大小、G分级、肝转移相关，TOPK阳性表达提示预后不良。本研究揭示了TOPK在胰腺神经内分泌肿瘤中的作用机制和肝转移的相关性，有重要的临床价值。