棕榈酸刺激的巨噬细胞对HepG2细胞侵袭和迁移的影响

目的:研究棕榈酸刺激的巨噬细胞对肝癌细胞侵袭和迁移能力的影响,并进一步探讨其作用机制。方法:应用棕榈酸(0.16 mmol/L)刺激人急性单核细胞白血病细胞系THP-1来源的巨噬细胞,并取其上清液处理HepG2细胞。细胞迁移实验检测巨噬细胞的迁移能力,侵袭实验和划痕实验分别观察HepG2细胞的纵向迁移能力和横向迁移能力;RT-qPCR检测巨噬细胞和HepG2细胞的炎症/趋化因子及HepG2细胞上皮-间充质转化标志蛋白(E-cadherin和N-cadherin)的mRNA表达水平。结果:棕榈酸促进了巨噬细胞迁移,显著上调巨噬细胞白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)和单核细胞趋化蛋白1(MCP-1)的mRNA表达;用棕榈酸刺激巨噬细胞的上清液处理HepG2细胞,其纵向迁移能力和横向迁移力明显强于未经棕榈酸处理组,且HepG2细胞的多种炎症因子和N-cadherin表达上调,E-cadherin表达下调。结论:棕榈酸可以增强巨噬细胞的迁移能力,刺激巨噬细胞产生大量炎症因子/趋化因子,进一步通过旁分泌/内分泌作用于HepG2细胞,促进HepG2细胞的上皮-间充质转化,增强HepG2细胞的侵袭与迁移能力。

Effects of palmitate-stimulated macrophages on invasion and migration of HepG2 cells

AIM: To investigate the impact of palmitate-stimulated macrophages on the invasion and migration of HepG2 cells and to explore the underlying mechanism. METHODS: Human acute monocytic leukemia cell line THP-1 were induced to macrophages by phorbol myristate acetate and were stimulated with palmitate (0.16 mmol/L). The culture supernatants were collected and used to incubate HepG2 cells. The effect of palmitate on migration of the macrophages was detected by Transwell chamber assay. The mRNA expression of target genes was measured by RT-qPCR. The invasion and migration of the HepG2 cells were assessed by invasion assay and scratch test. RESULTS: Palmitate promoted the migration of the macrophages and increased the mRNA levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and monocyte chemotactic protein-1 (MCP-1) in the macrophages. The invasion and migration of the HepG2 cells incubated with conditioned media from palmitate-stimulated macrophages were greater than those of the HepG2 cells incubated with conditioned media from macrophages without palmitate. The media of palmitate-stimulated macrophages up-regulated the mRNA expression of cytokines and N-cadherin, and down-regulated the mRNA expression of E-cadherin in the HepG2 cells. CONCLUSION: Palmitate-stimulated macrophages promote the invasion and migration of HepG2 cells through paracrine/endcrine loop.