PO.CL05.09 · 临床研究
热休克蛋白抑制剂以JAK、STAT依赖的方式抑制人胰腺癌细胞中细胞因子诱导的DUOX2 mRNA和蛋白表达
Heat shock protein inhibitors suppress cytokine-induced DUOX2 mRNA and protein expression in human pancreatic cancer cells in a JAK, STAT dependent manner
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
双氧化酶2(DUOX2)是七个NADPH氧化酶家族成员之一，在宿主防御和胃肠系统慢性炎症相关癌症中均发挥关键作用。在体外，促炎细胞因子如IFN-γ、IFNα/β、IL-4和IL-17A通过激活STATs和NF-κB信号通路蛋白增强DUOX2/DUOXA2的表达；在体内，与组织学正常的胰腺组织相比，DUOX蛋白和mRNA水平在慢性胰腺炎、癌前胰腺上皮内瘤变以及胰腺癌患者的早期阶段显著上调。在胰腺腺癌中，DUOX2表达升高与患者总生存率呈负相关。热休克蛋白90(Hsp90)是一种参与癌细胞肿瘤发生、侵袭和转移的重要分子伴侣，对许多致癌客户蛋白的折叠、成熟和稳定性至关重要，这些客户蛋白包括AKT和JAK1/2等激酶以及STAT3和HIF-1α等转录因子。若干STAT家族成员连同Hsp90在人胰腺癌中过表达。利用一组人胰腺癌细胞系(BxPC-3、AsPC-1和CFPAC-1)，我们发现两种不同的Hsp90抑制剂——坦螺旋霉素(17-AAG)和Ganetespib(STA-9090)——抑制JAK1和JAK2激酶，阻断细胞因子诱导的、JAK调控的STAT磷酸化。此外，这些Hsp90抑制剂以不同的敏感性抑制人胰腺癌细胞系中细胞因子诱导的DUOX2、VEGF-A、MMP-7和PD-L1表达。此外，JAK1/2抑制剂Ruxolitinib抑制BxPC-3细胞中IL-4诱导的、JAK介导的STAT6磷酸化以及DUOX2 mRNA和蛋白表达。使用JAK1、STAT1、2和STAT6特异性siRNA敲低也观察到类似结果。然而，用特异性siRNA同时敲低Hsp90的两种异构体Hsp90α和Hsp90β并未抑制JAK1活性、细胞因子诱导的DUOX2 mRNA或蛋白表达。细胞中残留的Hsp90蛋白或Hsp90的其他异构体可能会在siRNA敲低后代偿降低的Hsp90功能。我们的数据表明，Hsp90抑制剂通过阻断细胞因子激活的JAK-STATs致癌信号通路及其下游基因(如DUOX2、VEGF-A、MMP-7和PD-L1的表达)，可能成为炎症相关胰腺癌一种有价值的治疗方法。
查看英文原文 English abstract
Dual oxidase 2 (DUOX2), one of the seven NADPH oxidases family members, plays a critical role in both host defense and chronic inflammation-associated cancer in the gastrointestinal system. In vitro, pro-inflammatory cytokines, such as IFN- y, IFNalpha/beta, IL-4 and IL-17A, enhance DUOX2/DUOXA2 expression through activation of STATs and NF-kB signaling pathway proteins; in vivo, DUOX protein and mRNA levels are substantially upregulated in chronic pancreatitis, pre-malignant pancreatic intraepithelial neoplasms and the early stages of pancreatic cancer patients compared to histologically normal pancreatic tissues. In pancreatic adenocarcinoma, increased DUOX2 expression is adversely correlated with overall patient survival. Heat-Shock Protein 90 (Hsp90), an important molecular chaperone involved in tumorigenesis, invasion and metastasis of cancer cells, is critical in folding, maturation and stability of many oncogenic client proteins, including kinases such as AKT and JAK1/2, and transcription factors, such as STAT3 and HIF-1alpha. Several STAT family members, along with Hsp90, are overexpressed in human pancreatic carcinomas. Using a panel of human pancreatic cancer cell lines (BxPC-3, AsPC-1 and CFPAC-1), we found that two different Hsp90 inhibitors, Tanespimycin (17-AAG) and Ganetespib (STA-9090), inhibit JAK1 and JAK2 kinases, blocking cytokine-induced, JAK-regulated STAT phosphorylation. Additionally, these Hsp90 inhibitors suppress cytokine-induced DUOX2, VEGF-A, MMP-7 and PD-L1 expression in human pancreatic cancer cell lines with varying sensitivity. Furthermore, the JAK1/2 inhibitor Ruxolitinib inhibits IL-4 induced and JAK-mediated STAT6 phosphorylation, and DUOX2 mRNA and protein expression in BxPC-3 cells. Similar results were observed with JAK1, STAT1, 2 and STAT6 specific siRNA knockdown. However, simultaneous knockdown of both isoforms of Hsp90, Hsp90alpha and Hsp90beta, with specific siRNA did not inhibit JAK1 activity, cytokine-induced DUOX2 mRNA or protein expression. Either remaining Hsp90 protein or other isoforms of Hsp90 in cells may compensate decreased Hsp90 function after siRNA knockdown. Our data suggests that Hsp90 inhibitors, through blocking the cytokine-activated JAK-STATs oncogenic signaling pathway and their downstream genes such as DUOX2, VEGF-A, MMP-7 and PD-L1expression, may be a valuable therapeutic approach for inflammation-associated pancreatic cancer.
利益披露 Disclosure
Y. Wu, None..
A. Di, None..
S. Antony, None..
J. Meitzler, None..
M. M. Konaté, None..
K. Roy, None.