ABSTRACT

The aim of this study was to investigate the role of tumor-associated neutrophils (TANs) in the metastasis of pancreatic ductal adenocarcinoma (PDAC), to explore the regulation of TANs, and to determine the mechanisms governing the metastasis of PDAC. The correlation between neutrophils and the patient’s clinical pathological data was first evaluated. Then, the effects of neutrophils on the invasion of PDAC were analyzed using a combination of conditioned media, direct and indirect coculture of human peripheral blood neutrophils, and PDAC cell lines (Panc-1, MiaPaCa-2 and AsPC-1). The cytokines secreted by neutrophils were detected through ELISA. TAN density was significantly correlated with poor metastasis-free survival (P < .05). Through coculture, it was found that the effect of neutrophils on pancreatic cancer cells was dependent on concentration, and a high concentration of neutrophils showed a lethal effect, while a low concentration of neutrophils primarily promoted the migration ability of cancer cells. The results of the wound-healing assay, the Transwell invasion assay, and laser confocal microscopy confirmed the promoting effect and indicated that the effect of neutrophils toward cancer cells may function indirectly by releasing a series of cytokines. The results of ELISA show that this effect may be achieved through the secretion of a large amount of TNF-α and TGF-β1 by neutrophils. Our study indicated that neutrophils may increase the metastasis of PDAC by releasing a series of cell cytokines, such as TNF-α and TGF-β1.

摘要

本研究旨在探讨肿瘤相关中性粒细胞（TANs）在胰腺导管腺癌（PDAC）转移中的作用，探讨TANs的调控作用，并确定控制PDAC转移的机制。首先评估中性粒细胞与患者临床病理数据之间的相关性。然后，使用条件培养基、人外周血中性粒细胞的直接和间接共培养和 PDAC 细胞系（Panc-1、MiaPaCa-2 和 AsPC-1）的组合分析中性粒细胞对 PDAC 侵袭的影响。通过ELISA检测中性粒细胞分泌的细胞因子。TAN 密度与较差的无转移生存期显着相关（P< .05)。通过共培养发现中性粒细胞对胰腺癌细胞的作用依赖于浓度，高浓度的中性粒细胞表现出致死作用，而低浓度的中性粒细胞主要促进癌细胞的迁移能力。伤口愈合试验、Transwell 侵袭试验和激光共聚焦显微镜的结果证实了促进作用，并表明中性粒细胞对癌细胞的作用可能通过释放一系列细胞因子间接发挥作用。ELISA结果表明，这种作用可能是通过中性粒细胞分泌大量的TNF-α和TGF-β1来实现的。我们的研究表明，中性粒细胞可能通过释放一系列细胞因子，如 TNF-α 和 TGF-β1 来增加 PDAC 的转移。