ANXA1通过PKC/JNK/P-gp通路调控胰腺癌原发耐药的分子机制研究

De novo resistance is the leading cause of death in pancreatic cancer patients. In our prophase research, we have found that low expression of ANXA1 is involved in the regulation of de novo resistance of pancreatic cancer. In our pre experiment, we have found that low expression of ANXA1 can reduce the drug sensitivity of pancreatic cancer cells. Because ANXA1 can bind to PKC and inhibit the function of PKC, in our further testing, we found the proteins PKC, JNK and P-gp belong to PKC pathway were upregulated. Consequently, we suppose that ANXA1 may regulate the drug resistance of pancreatic cancer through PKC/JNK/P-gp pathway. .The projects include: (1) We will study the molecular mechanism of ANXA1 regulates the de novo resistance of pancreatic cancer through PKC/JNK/P-gp pathway by in vitro cell model; (2) We will verify whether ANXA1 can regulate de novo drug resistance through PKC/JNK/P-gp pathway by in vivo nude mouse model; (3) We will analysis the correlation between the ANXA1, PKC, JNK and P-gp expresson and the pathology and prognosis of pancreatic cancer patients by 196 cases tissue microarray. Our study will show that ANXA1 may be an important marker for human pancreatic cancer drug resistance and prognosis as well as a potential therapeutic target.

原发耐药是胰腺癌患者死亡的主要原因。申请者利用已构建的细胞模型通过蛋白质组学方法研究发现ANXA1低表达与胰腺癌原发耐药有关。申请者预实验发现下调ANXA1表达能够降低胰腺癌细胞的药物敏感性。因ANXA1可以与PKC结合并抑制PKC的功能，申请者进一步检测发现下调ANXA1表达后，PKC/JNK/P-gp耐药通路蛋白表达均上调。据此，本课题提出假说：ANXA1通过调控PKC/JNK/P-gp通路调控胰腺癌耐药。.为此，我们将（1）构建ANXA1高、低表达细胞模型，研究ANXA1在胰腺癌原发耐药中的作用及其调控PKC/JNK/P-gp通路的机制；（2）用裸鼠模型验证ANXA1在胰腺癌原发耐药中的作用及其对PKC/JNK/P-gp通路的影响;（3）用196例胰腺癌组织芯片分析ANXA1与PKC、JNK和P-gp表达之间的关系，以及与胰腺癌预后的关系。本研究将为寻找胰腺癌治疗靶点提供理论依据。

胰腺癌临床表现隐匿，确诊时80%的患者失去手术机会，所以化疗是胰腺临床治疗的重要手段，但其对多种化疗药物均不敏感，原发耐药是胰腺癌患者死亡的主要原因。本课题在前期已构建的胰腺癌原发耐药细胞模型基础上通过蛋白质组学方法研究筛选出与胰腺癌原发耐药有关的蛋白质ANXA1，进而在体外细胞及体内裸鼠模型两方面通过western blot方法进一步验证蛋白质组学的结果，结果与蛋白质组学的结果一致，证实ANXA1与胰腺癌耐药有关。选择两株胰腺癌细胞株通过siRNA构建低表达ANXA1的胰腺癌细胞株后，通过药物敏感性实验研究证实下调ANXA1的表达能降低胰腺癌细胞的药物敏感性。进一步通过western blot方法分析下调ANXA1的表达后，PKC, p-PKC, JNK, p-JNK和P-gp的表达量明显增高。综合上述结果，本研究证实ANXA1通过PKC/JNK/P-gp通路调控胰腺癌原发耐药，为临床胰腺癌治疗靶点的筛选提供理论依据。