Department of Surgery, Helsinki University Central Hospital, University of Helsinki, Post Box 263, 00029, Helsinki, Finland.
Med Oncol. 2014 Mar;31(3):884. doi: 10.1007/s12032-014-0884-9. Epub 2014 Feb 14.
Patients with chronic pancreatitis with local inflammation have high risk for pancreatic cancer. The aim of this study was to examine the role of the inflammatory cells in the invasion of pancreatic cancer cells, focusing on the involvement of a disintegrin and metalloproteinase 8 (ADAM8) and matrix metalloproteinase 9 (MMP9) proteins. ADAM8 expression is associated with worse survival of pancreatic cancer patients. Monocytes from healthy donors were differentiated into macrophages. Pancreatic adenocarcinoma cells were cultured either alone or with differentiated macrophages. The cancer cell migration rate in Matrigel was measured by imaging fluorescently stained cells for 24 h. After invasion, cells were sorted into CD14 positive/negative macrophages and cancer cells with magnetic separation. The expression of ADAM8 and MMP9 was measured by the real-time PCR. Protein-level expression of ADAM8 and MMP9 was analyzed by Western blotting. In two series, siRNA technique was used to reduce either ADAM8 or MMP9 expression in the cancer cells. The coculture with macrophages increased cancer cell migration rate in Matrigel, and increased ADAM8 and MMP9 mRNA expression and protein level in the cancer cells. Reduction of ADAM8 expression with siRNA in the cancer cells decreased macrophage-induced migration rate of the cancer cells from 11.7±0.3 μm/h to 9.0±0.2 μm/h (p<0.01), and reduction of MMP9 expression decreased the migration rate to 10.1±0.2 μm/h (p<0.01). Anti-inflammatory macrophages increase pancreatic cancer cell migration rate in basement membrane matrix by inducing ADAM8 and MMP9 expression in cancer cells, thereby possibly enhancing the invasiveness of cancer.
患有局部炎症的慢性胰腺炎患者患胰腺癌的风险较高。本研究旨在研究炎症细胞在胰腺癌细胞浸润中的作用，重点研究解整合素金属蛋白酶 8 (ADAM8) 和基质金属蛋白酶 9 (MMP9) 蛋白的参与。ADAM8 的表达与胰腺癌患者的生存预后较差相关。从健康供体中分离出单核细胞并分化为巨噬细胞。将胰腺腺癌细胞单独培养或与分化的巨噬细胞共培养。通过对 24 小时荧光染色的细胞进行成像来测量癌细胞在 Matrigel 中的迁移率。侵袭后，通过磁性分离将细胞分为 CD14 阳性/阴性巨噬细胞和癌细胞。通过实时 PCR 测量 ADAM8 和 MMP9 的表达。通过 Western blot 分析 ADAM8 和 MMP9 的蛋白水平表达。在两个系列中，使用 siRNA 技术降低癌细胞中 ADAM8 或 MMP9 的表达。与巨噬细胞共培养可增加癌细胞在 Matrigel 中的迁移率，并增加癌细胞中 ADAM8 和 MMP9 的 mRNA 表达和蛋白水平。用 siRNA 降低癌细胞中的 ADAM8 表达可将巨噬细胞诱导的癌细胞迁移率从 11.7±0.3 μm/h 降低至 9.0±0.2 μm/h (p<0.01)，降低 MMP9 表达可将迁移率降低至 10.1±0.2 μm/h (p<0.01)。抗炎性巨噬细胞通过诱导癌细胞中 ADAM8 和 MMP9 的表达来增加胰腺癌细胞在基底膜基质中的迁移率，从而可能增强癌症的侵袭性。
