Kanemaru Mikio, Maehara Naoki, Iwamura Takeshi, Chijiiwa Kazuo
Department of Surgical Oncology and Regulation of Organ Function, Miyazaki University School of Medicine, Miyazaki, Japan.
Hepatogastroenterology. 2012 Jul-Aug;59(117):1614-20. doi: 10.5754/hge10036.
BACKGROUND/AIMS: The aim of this study was to investigate the effect of thrombin and the thrombin receptor protease-activated receptor (PAR)-1 on adhesion of human pancreatic cancer cell lines to extracellular matrices (ECMs) and to identify related integrins with these effects.
Human pancreatic cancer cell lines SUIT-2 and its four sublines, and Panc- 1, AsPC-1 and MiaPaCa-2 were treated with thrombin, PAR-1 agonist TRAP-6, PAR-1 antagonist SCH79797, or anti-integrin ±vβ3, ±vβ5 and β1 monoclonal antibodies. Cells were incubated for 45 minutes on micro titer plates that were pre-coated with ECMs (fibronectin, laminin, vitronectin, type IV collagen). The number of adherent cells was measured by the MTT method.
Eight human pancreatic cancer cell lines expressed PAR-1. Thrombin significantly enhanced adhesion of SUIT-2 and its sublines and MiaPaCa-2 to vitronectin, especially in the SUIT-2 subline S2-007. We obtained similar results on S2-007 cells through treatment with TRAP-6. However, SCH79797 inhibited the effect of thrombin. Furthermore, anti-integrin β1 antibody conspicuously inhibited 1U/mL thrombin-induced enhancement of adhesion to vitronectin.
Thrombin significantly enhanced adhesion of pancreatic cancer cells to vitronectin through PAR- 1 depending on the presence of integrin β1. Suppression of thrombin action by anti-integrin β1 antibody will become a useful therapy against pancreatic cancer.
背景/目的：本研究旨在探讨凝血酶及凝血酶受体蛋白酶激活受体（PAR）-1对人胰腺癌细胞系与细胞外基质（ECM）黏附的影响，并确定与这些作用相关的整合素。
用人胰腺癌细胞系SUIT-2及其四个亚系，以及Panc-1、AsPC-1和MiaPaCa-2细胞，分别用凝血酶、PAR-1激动剂TRAP-6、PAR-1拮抗剂SCH79797或抗整合素±vβ3、±vβ5和β1单克隆抗体处理。将细胞在预先包被有ECM（纤连蛋白、层粘连蛋白、玻连蛋白、IV型胶原）的微量滴定板上孵育45分钟。采用MTT法测定黏附细胞数。
8种人胰腺癌细胞系表达PAR-1。凝血酶显著增强SUIT-2及其亚系和MiaPaCa-2细胞对玻连蛋白的黏附，尤其是在SUIT-2亚系S2-007中。用TRAP-6处理S2-007细胞也得到了类似结果。然而，SCH79797抑制了凝血酶的作用。此外，抗整合素β1抗体显著抑制1U/mL凝血酶诱导的对玻连蛋白黏附增强。
凝血酶通过PAR-1显著增强胰腺癌细胞对玻连蛋白的黏附，这依赖于整合素β1的存在。抗整合素β1抗体抑制凝血酶作用将成为一种有效的胰腺癌治疗方法。
