粘着斑激酶的激活通过细胞外信号调节激酶-1/2信号通路的激活增强胰腺癌细胞的粘附和侵袭。
Activation of focal adhesion kinase enhances the adhesion and invasion of pancreatic cancer cells via extracellular signal-regulated kinase-1/2 signaling pathway activation.
作者信息
Sawai Hirozumi, Okada Yuji, Funahashi Hitoshi, Matsuo Yoichi, Takahashi Hiroki, Takeyama Hiromitsu, Manabe Tadao
机构信息
Department of Gastroenterological Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya 4678601, Japan.
出版信息
Mol Cancer. 2005 Oct 6;4:37. doi: 10.1186/1476-4598-4-37.
BACKGROUND
Interaction with integrin and focal adhesion kinase (FAK) regulates the cancer cell adhesion and invasion into extracellular matrix (ECM). In addition, phosphorylation of FAK correlates with the increase of cell motility and invasion. Adhesion and spreading of cancer cells on a variety of ECM proteins, including collagen type IV (Coll IV), leads to an increase in tyrosine phosphorylation and activation of FAK. In this study, we investigated the mechanism of activation of FAK and its downstream extracellular signal-regulated kinase (ERK)-1/2 signaling following stimulation by interleukin (IL)-1alpha and adhesion to ECM with subsequent enhancement of pancreatic cancer cell adhesion and invasion.
RESULTS
In immunoblotting analysis, all three pancreatic cancer cell lines (AsPC-1, BxPC-3, and Capan-2) expressed the protein of FAK and beta1 integrin. Enhancement of FAK protein association with beta1 integrin when cells were plated on Coll IV was more increased by stimulation with IL-1alpha. Preincubation with anti-beta1 integrin antibody and FAK siRNA transfection inhibited the association of FAK with beta1 integrin of pancreatic cancer cells. FAK phosphorylation was observed by adhesion to Coll IV, furthermore, stronger FAK phosphorylation was observed by stimulation with IL-1alpha of pancreatic cancer cells adhered to Coll IV in time-dependent manner. Genistein, a tyrosine kinase inhibitor, markedly inhibited the FAK phosphorylation. IL-1alpha stimulation and Coll IV adhesion enhanced the activation of Ras, as evidenced by the increased Ras-GTP levels in pancreatic cancer cells. Activation of Ras correlated with the phosphorylation of ERK. While not statistical affecting the apoptosis of pancreatic cancer cells, IL-1alpha-induced adhesion and invasion on Coll IV were inhibited with FAK gene silencing by siRNA, beta1 integrin blocking, and inhibition of FAK phosphorylation. PD98059, a MEK inhibitor, also inhibited IL-1alpha-induced enhancement of adhesion and invasion in pancreatic cancer cells.
CONCLUSION
Our results demonstrated that activation of FAK is involved with the aggressive capability in pancreatic cancer through Ras/ERK signaling pathway. Based on our results, we suggest that the modification of IL-1, FAK, and integrins functions might be a novel therapeutic approach to aggressive spread of pancreatic cancer.
背景
与整合素和粘着斑激酶（FAK）的相互作用调节癌细胞对细胞外基质（ECM）的粘附和侵袭。此外，FAK的磷酸化与细胞运动性和侵袭性的增加相关。癌细胞在包括IV型胶原（Coll IV）在内的多种ECM蛋白上的粘附和铺展，会导致酪氨酸磷酸化增加以及FAK的激活。在本研究中，我们调查了白细胞介素（IL）-1α刺激以及与ECM粘附后FAK及其下游细胞外信号调节激酶（ERK）-1/2信号通路激活的机制，随后胰腺癌细胞的粘附和侵袭增强。
结果
在免疫印迹分析中，所有三种胰腺癌细胞系（AsPC-1、BxPC-3和Capan-2）均表达FAK和β1整合素蛋白。当细胞接种在Coll IV上时，IL-1α刺激使FAK蛋白与β1整合素的结合增强。用抗β1整合素抗体预孵育和FAK siRNA转染抑制了胰腺癌细胞中FAK与β1整合素的结合。通过粘附到Coll IV观察到FAK磷酸化，此外，通过IL-1α刺激粘附到Coll IV的胰腺癌细胞，以时间依赖性方式观察到更强的FAK磷酸化。酪氨酸激酶抑制剂染料木黄酮显著抑制FAK磷酸化。IL-1α刺激和Coll IV粘附增强了Ras的激活，胰腺癌细胞中Ras-GTP水平升高证明了这一点。Ras的激活与ERK的磷酸化相关。虽然对胰腺癌细胞的凋亡没有统计学影响，但通过siRNA使FAK基因沉默、β1整合素阻断和抑制FAK磷酸化，抑制了IL-1α诱导的在Coll IV上的粘附和侵袭。MEK抑制剂PD98059也抑制了IL-1α诱导的胰腺癌细胞粘附和侵袭增强。
结论
我们的结果表明，FAK的激活通过Ras/ERK信号通路参与胰腺癌的侵袭能力。基于我们的结果，我们建议改变IL-1、FAK和整合素的功能可能是一种针对胰腺癌侵袭性扩散的新型治疗方法。
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