靶向干预糖原合成酶激酶-3β抑制胰腺癌血管生成、侵袭及转移的分子机制研究

GSK-3β is regarded as the attractive target in cancer therapy since many researches have showed that the inhibition of GSK-3β could inhibit the proliferation and induce the apoptosis of cancer cells.Very recently, we have found that the inhibition of GSK-3β inhibits the angiogenesis of pancreatic cancer, while the molecular mechanisns are largely unknown. Since NOTCH signal pathway plays the important role in the angiogenesis of cancer, again our prelimilary data showed that GSK-3β could regulate NOTCH molecuar specifically. We speculate that trageted GSK-3β may inhibit angiogenesis, invasion and metastasis of pancreatic cancer through NOTCH signal pathway. Here,Using immunohistochemistry, co-IP,immmuno-image in vivo, et al., we try to understand the mechenism of GSK-3β regulating NOTCH signal pathway including the ways and the location of interreaction in cancer cells , and to address the effects of GSK-3β on the proliferation,survival,migration and invasion in cancer cells,vascular endothenial cells and vascular smooth muscle cells, and also to assess whether GSK-3β plays the role depending on NOTCH signal pathway or not. Through this project, hopefully we would uncover the molecular mechanisms of GSK-3β having effects on angiogenises,invasion and metastasis in pancreatic cance, and also get the supportive evidences for the clinical usage of small molecular GSK-3β inhibitor.

最近多项研究显示靶向干预糖原合成酶激酶-3β（GSK-3β）可抑制癌细胞增殖、诱导凋亡，认为是胰腺癌靶向治疗极具潜力的靶点。我们新近发现GSK-3β可调控胰腺癌新生血管形成，但分子机制尚不清楚。由于NOTCH信号通路在肿瘤血管生成中发挥重要作用，且预实验结果提示GSK-3β特异调节NOTCH3分子，我们推测靶向抑制GSK-3β可能通过NOTCH信号通路抑制胰腺癌血管生成、侵袭和转移。为此，本项目拟从组织、细胞及整体三个水平，采用组化、免疫共沉淀、活体荧光成像等方法，研究GSK-3β调控NOTCH3的作用方式、定位及机制，探索GSK-3β对胰腺癌细胞、血管内皮细胞、血管平滑肌细胞生物学行为影响及分子机制，评价GSK-3β发挥作用是否依赖NOTCH信号通路。通过完成本项目，明确GSK-3β调控胰腺癌血管生成、侵袭及转移的分子机制，为干预GSK-3β小分子靶向药物临床应用奠定理论基础。

最近多项研究显示靶向干预糖原合成酶激酶-3β（GSK-3β）可抑制癌细胞增殖、诱导凋亡，而且 GSK-3β可调控胰腺癌新生血管形成，由于 NOTCH 信号通路在肿瘤血管生成中发挥重要作用，因此我们推测靶向抑制 GSK-3β可能通过 NOTCH 信号通路抑制胰腺癌血管生成、侵袭和转移。首先在胰腺癌临床标本中我们发现GSK-3β及NOTCH3在胰腺癌组织中表达显著高于癌旁组织，而且GSK-3β和NOTCH3的表达呈正相关，同时我们发现GSK-3β表达与胰腺癌临床分期，TNM分期、肿瘤分化程度及淋巴结转移之间存在正相关（p<0.05），而与患者年龄、性别及肿瘤大小之间无相关性（p>0.05）。最后，建立NOTCH3Q敲出胰腺癌细胞模型体内外探讨靶向干预NOTCH3对胰腺癌增殖侵袭及血管生成等影响。通过完 成本项目，明确了GSK-3β调控胰腺癌血管生成、侵袭及转移的分子机制，为干预 GSK-3β小分子靶向药物临床应用奠定了理论基础。