Center for Cancer Research, National Cancer Institute, Bethesda, MD 20892, USA.
J Mol Neurosci. 2012 Jan;46(1):68-74. doi: 10.1007/s12031-011-9639-7. Epub 2011 Sep 6.
The effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on tyrosine phosphorylation of focal adhesion kinase (FAK) and paxillin were investigated using lung cancer cells. Addition of PACAP-27 or PACAP-38 but not vasoactive intestinal peptide to NCI-H838 or NCI-H1299 human lung cancer cells significantly increased the tyrosine phosphorylation of FAK or paxillin. The increase in FAK or paxillin tyrosine phosphorylation caused by addition of PACAP-27 to NCI-H838 cells was inhibited by PACAP(6-38), a PAC1-receptor (R) antagonist. The increase in FAK or paxillin tyrosine phosphorylation caused by 100 nM PACAP-27 was maximal 2 min after addition to NCI-H838 cells. The effects of PACAP at stimulating FAK and paxillin tyrosine phosphorylation were reversed by cytochalasin D and genistein which inhibit actin polymerization and tyrosine kinase activity, respectively. The effects of PACAP at stimulating FAK and paxillin tyrosine phosphorylation were reversed by U-73122 but not H89 which inhibit phospholipase C and protein kinase A, respectively. The results show that PAC1-R regulates FAK and paxillin tyrosine phosphorylation in lung cancer cells as a result of increased phosphatidylinositol turnover but not adenylyl cylase stimulation.
研究了垂体腺苷酸环化酶激活肽（PACAP）对肺癌细胞中粘着斑激酶（FAK）和桩蛋白酪氨酸磷酸化的影响。将 PACAP-27 或 PACAP-38 而不是血管活性肠肽添加到 NCI-H838 或 NCI-H1299 人肺癌细胞中，可显著增加 FAK 或桩蛋白的酪氨酸磷酸化。PACAP(6-38)，一种 PAC1 受体（R）拮抗剂，抑制 PACAP-27 对 NCI-H838 细胞中 FAK 或桩蛋白酪氨酸磷酸化的增加。PACAP-27 对 NCI-H838 细胞的刺激作用在添加 2 分钟后达到最大，100 nM PACAP-27 可使 FAK 或桩蛋白酪氨酸磷酸化增加。细胞松弛素 D 和金雀异黄素分别抑制肌动蛋白聚合和酪氨酸激酶活性，可逆转 PACAP 刺激 FAK 和桩蛋白酪氨酸磷酸化的作用。U-73122 可逆转 PACAP 刺激 FAK 和桩蛋白酪氨酸磷酸化的作用，但 H89 不能，H89 抑制磷脂酶 C 和蛋白激酶 A。结果表明，PAC1-R 通过增加磷酸肌醇周转率而不是刺激腺苷酸环化酶调节肺癌细胞中 FAK 和桩蛋白的酪氨酸磷酸化。
