Insulin Receptor and GPCR Crosstalk Stimulates YAP via PI3K and PKD in Pancreatic Cancer Cells.
Insulin Receptor and GPCR Crosstalk Stimulates YAP via PI3K and PKD in Pancreatic Cancer Cells.
复制标题
DOI：
10.1158/1541-7786.mcr-17-0023
复制
发表时间：
2017-07
期刊：
影响因子：
--
通讯作者：
Rozengurt E
中科院分区：
文献类型：
--
作者：
Hao F;Xu Q;Zhao Y;Stevens JV;Young SH;Sinnett-Smith J;Rozengurt E
我们研究了胰岛素受体（INSR）和G蛋白偶联受体（GPCR）信号通路之间的串扰对人胰腺导管腺癌（PDAC）中Yes相关蛋白（雅普）定位、磷酸化和转录活性的调节的影响。用胰岛素和神经降压素（这些细胞的有效促有丝分裂激动剂组合）刺激PANC-1或MiaPaCa-2细胞，促进了显著的雅普核定位，并降低了雅普在Ser 127和Ser 397的磷酸化。单独用胰岛素或神经降压素刺激PDAC细胞适度诱导雅普/TEAD调节基因的表达，包括结缔组织生长因子（CTGF）、富含半胱氨酸的血管生成诱导物61（CYR 61）和CXCL 5，而神经降压素和胰岛素的组合诱导这些基因的表达水平显著增加。此外，siRNA介导的雅普/TAZ敲低阻止了这些基因表达的增加。一种对PI 3 K的p110α亚基具有选择性的小分子抑制剂（A66）消除了神经降压素和胰岛素诱导的3，4，5-三磷酸磷脂酰肌醇（PIP 3）产生以及CTGF、CYR 61和CXCL 5表达的增加。此外，用蛋白激酶D（PKD）家族抑制剂（CRT 0066101或kb NB 142-70）或用靶向PKD家族的siRNA处理PDAC细胞防止了响应于胰岛素和神经降压素刺激的CTGF、CYR 61和CXCL 5 mRNA水平的增加。因此，在胰腺癌细胞中，PI 3 K和PKD介导响应于胰岛素和神经降压素的雅普活化。
We examined the impact of crosstalk between the insulin receptor (INSR) and G protein-coupled receptor (GPCR) signaling pathways on the regulation of Yes-associated Protein (YAP) localization, phosphorylation and transcriptional activity in the context of human pancreatic ductal adenocarcinoma (PDAC). Stimulation of PANC-1 or MiaPaCa-2 cells with insulin and neurotensin, a potent mitogenic combination of agonists for these cells, promoted striking YAP nuclear localization and decreased YAP phosphorylation at Ser127 and Ser397. Challenging PDAC cells with either insulin or neurotensin alone modestly induced the expression of YAP/TEAD-regulated genes, including Connective Tissue Growth Factor (CTGF), Cysteine-rich angiogenic inducer 61 (CYR61) and CXCL5 whereas the combination of neurotensin and insulin induced a marked increase in the level of expression of these genes. In addition, siRNA-mediated knockdown of YAP/TAZ prevented the increase in the expression of these genes. A small-molecule inhibitor (A66), selective for the p110α subunit of PI3K, abrogated the increase in phosphatidylinositol 3,4,5-trisphosphate (PIP3) production and the expression of CTGF, CYR61 and CXCL5 induced by neurotensin and insulin. Furthermore, treatment of PDAC cells with protein kinase D (PKD) family inhibitors (CRT0066101 or kb NB 142-70) or with siRNAs targeting the PKD family prevented the increase of CTGF, CYR61 and CXCL5 mRNA levels in response to insulin and neurotensin stimulation. Thus, PI3K and PKD mediate YAP activation in response to insulin and neurotensin in pancreatic cancer cells.