Cyclic GMP/protein kinase G type-Iα (PKG-Iα) signaling pathway promotes CREB phosphorylation and maintains higher c-IAP1, livin, survivin, and Mcl-1 expression and the inhibition of PKG-Iα kinase activity synergizes with cisplatin in non-small cell lung cancer cells
Cyclic GMP/protein kinase G type-Iα (PKG-Iα) signaling pathway promotes CREB phosphorylation and maintains higher c-IAP1, livin, survivin, and Mcl-1 expression and the inhibition of PKG-Iα kinase activity synergizes with cisplatin in non-small cell lung cancer cells
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DOI：
10.1002/jcb.24237
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发表时间：
2012-11-01
影响因子：
4
通讯作者：
Fiscus, Ronald R.
中科院分区：
文献类型：
--
作者：
Wong, Janica C.;Bathina, Madhavi;Fiscus, Ronald R.
此前，我们实验室发现一氧化氮（NO）/环鸟苷酸（cGMP）/蛋白激酶G Ia型（PKG-Ia）信号通路在正常细胞（骨髓基质细胞和血管平滑肌细胞）和某些癌细胞（卵巢癌细胞）中，在防止自发性凋亡和促进细胞增殖方面发挥着重要作用。在本研究中，我们研究了cGMP/PKG-Ia通路在NCI-H460和A549非小细胞肺癌（NSCLC）细胞中预防自发性细胞凋亡、促进集落形成、调节cAMP反应元件结合（CREB）蛋白磷酸化以及凋亡抑制蛋白（IAP）和抗凋亡Bcl-2相关蛋白表达的新作用。 1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one (ODQ) 可阻断内源性 NO 诱导的 cGMP/PKG-Ia 激活，诱导细胞凋亡并减少集落形成。 ODQ 还降低 CREB ser133 磷酸化以及 c-IAP1、livin 和 survivin 的蛋白表达。 DT-2（PKG-Ia 激酶活性抑制剂）使细胞凋亡增加两倍，并降低 CREB ser133 磷酸化和 c-IAP1、livin 和 survivin 表达。使用小干扰 RNA 基因敲低 PKG-Ia 表达可增加细胞凋亡并减少 CREB ser133 磷酸化以及 c-IAP1、livin、survivin 和 Mcl-1 表达。用 DT-2 抑制 PKG-Ia 激酶活性可显着增强化疗药物顺铂的促凋亡作用。与单独使用顺铂或DT-2相比，DT-2和顺铂联合治疗可增加细胞凋亡，显示出协同作用。数据表明，PKG-Ia 激酶活性对于维持较高水平的 CREB ser133 磷酸化以及 c-IAP1、livin、survivin 和 Mcl-1 的蛋白表达是必需的，从而防止 NSCLC 细胞自发凋亡并促进集落形成，这可能会限制顺铂等化疗药物的有效性。 J.细胞。生物化学。 113：35873598，2012。(C)2012 Wiley 期刊公司。
Previously, our laboratory showed that nitric oxide (NO)/cyclic GMP (cGMP)/protein kinase G type-Ia (PKG-Ia) signaling pathway plays an important role in preventing spontaneous apoptosis and promoting cell proliferation in both normal cells (bone marrow stromal cells and vascular smooth muscle cells) and certain cancer cells (ovarian cancer cells). In the present study, we investigated the novel role of the cGMP/PKG-Ia pathway in preventing spontaneous apoptosis, promoting colony formation and regulating phosphorylation of cAMP response element binding (CREB) protein and protein expression of inhibitor of apoptosis proteins (IAPs) and anti-apoptotic Bcl-2-related proteins in NCI-H460 and A549 non-small cell lung cancer (NSCLC) cells. 1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one (ODQ), which blocks endogenous NO-induced activation of cGMP/PKG-Ia, induced apoptosis and decreased colony formation. ODQ also decreased CREB ser133 phosphorylation and protein expression of c-IAP1, livin, and survivin. DT-2 (inhibitor of PKG-Ia kinase activity) increased apoptosis by twofold and decreased CREB ser133 phosphorylation and c-IAP1, livin, and survivin expression. Gene knockdown of PKG-Ia expression using small-interfering RNA increased apoptosis and decreased CREB ser133 phosphorylation, and c-IAP1, livin, survivin, and Mcl-1 expression. Inhibition of PKG-Ia kinase activity with DT-2 dramatically enhanced pro-apoptotic effects of the chemotherapeutic agent cisplatin. Combined treatment of DT-2 and cisplatin increased apoptosis compared with cisplatin or DT-2 alone, showing a synergistic effect. The data suggest that the PKG-Ia kinase activity is necessary for maintaining higher levels of CREB phosphorylation at ser133 and protein expression of c-IAP1, livin, survivin, and Mcl-1, preventing spontaneous apoptosis and promoting colony formation in NSCLC cells, which may limit the effectiveness of chemotherapeutic agents like cisplatin. J. Cell. Biochem. 113: 35873598, 2012. (C) 2012 Wiley Periodicals, Inc.