Induced Overexpression of Protein Kinase D1 Stimulates Mitogenic Signaling in Human Pancreatic Carcinoma PANC-1 Cells
Induced Overexpression of Protein Kinase D1 Stimulates Mitogenic Signaling in Human Pancreatic Carcinoma PANC-1 Cells
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DOI：
10.1002/jcp.22036
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发表时间：
2010-05-01
影响因子：
5.6
通讯作者：
Rozengurt, Enrique
中科院分区：
文献类型：
--
作者：
Kisfalvi, Krisztina;Hurd, Cliff;Rozengurt, Enrique
神经紧张素（NT）刺激人胰腺腺癌细胞系PANC-1中蛋白激酶D1 （PKD1）、细胞外信号调节激酶（ERK）、c-Jun n -末端激酶（JNK）和DNA合成。为了确定PKD1过表达对这些生物反应的影响，我们产生了可诱导的稳定的PANC-1克隆，这些克隆表达野生型（WT）或激酶死亡（K618N）形式的PKD1，以响应脱皮激素类似物ponasterone-A （PonA）。在野生型和克隆型PANC-1细胞中，NT都能有效地刺激c-Jun Ser（63）的磷酸化。pona诱导的WT表达，而不是K618N PKD1，在MKK4水平或上游迅速阻断nt介导的c-Jun Ser（63）磷酸化，MKK4是一种导致JNK激活的双特异性激酶。这是PKD1抑制nt诱导的PANC-1细胞中JNK/cJun激活的首次证明。相比之下，PKD1过表达显著增加了nt诱导的ERK激活在这些细胞中的持续时间。PKD1信号传导对促有丝分裂erk和促凋亡JNK/c-Jun通路的相互影响促使我们研究PKD1过表达是否促进了PANC-1细胞的DNA合成和增殖。我们的研究结果表明，PKD1过表达增加了PANC-1细胞在常规培养皿或聚甲基丙烯酸羟乙酯（聚（HEMA））包被培养皿中培养的DNA合成和细胞数量，以消除细胞粘附（不依赖锚定生长）。PKD1过表达显著增强NT （1-10 nM）诱导的DNA合成。这些结果表明PKD1介导PANC-1中的有丝分裂信号传导，并表明该酶可能成为开发限制这些细胞增殖的治疗药物的新靶点。j .细胞。中国生物医学工程学报，2009,31(2):389 - 391。(c) 2010 Wiley-Liss, Inc。
Neurotensin (NT) stimulates protein kinase D1 (PKD1), extracellular signal regulated kinase (ERK), c-Jun N-terminal Kinase (JNK), and DNA synthesis in the human pancreatic adenocarcinoma cell line PANC-1. To determine the effect of PKD1 overexpression on these biological responses, we generated inducible stable PANC-1 clones that express wild-type (WT) or kinase-dead (K618N) forms of PKD1 in response to the ecdysone analog ponasterone-A (PonA). NT potently stimulated c-Jun Ser(63) phosphorylation in both wild type and clonal derivatives of PANC-1 cells. PonA-induced expression of WT, but not K618N PKD1, rapidly blocked NT-mediated c-Jun Ser(63) phosphorylation either at the level of or upstream of MKK4, a dual-specificity kinase that leads to JNK activation. This is the first demonstration that PKD1 suppresses NT-induced JNK/cJun activation in PANC-1 cells. In contrast, PKD1 overexpression markedly increased the duration of NT-induced ERK activation in these cells. The reciprocal influence of PKD1 signaling on pro-mitogenicERK and pro-apopotic JNK/c-Jun pathways prompted us to examine whether PKD1 overexpression promotes DNA synthesis and proliferation of PANC-1 cells. Our results show that PKD1 overexpression increased DNA synthesis and cell numbers of PANC-1 cells cultured in regular dishes or in polyhydroxyethylmethacrylate [Poly-(HEMA)]-coated dishes to eliminate cell adhesion (anchorage-independent growth). Furthermore, PKD1 overexpression markedly enhanced DNA synthesis induced by NT (1-10 nM). These results indicate that PKD1 mediates mitogenic signaling in PANC-1 and suggests that this enzyme could be a novel target for the development of therapeutic drugs that restrict the proliferation of these cells. J. Cell. Physiol. 223: 309-316, 2010. (c) 2010 Wiley-Liss, Inc.