Oral Biology Research Unit, Faculty of Dentistry, Thammasat University (Rangsit Campus), Pathum Thani, Thailand.
Biomed Pharmacother. 2018 May;101:988-995. doi: 10.1016/j.biopha.2018.03.033. Epub 2018 Mar 22.
The aim of this study was to investigate the effect of Porphyromonas gingivalis lipopolysaccharide (LPS)-induced macrophages on head and neck squamous cell carcinoma (HNSCC) cell line proliferation and invasion.
THP-1 monocytes were differentiated toward macrophages using 12.5 ng/ml phorbol 12-myristate 13-acetate treatment for 48 h. The expression of interleukin-6 (IL-6) mRNA and cytokine by monocytes and macrophages was determined using real time PCR and ELISA, respectively. The cells were analyzed for CD14 expression using immunofluorescent labeling. The macrophages were induced using 1 μg/ml P. gingivalis LPS for 24 h, and the conditioned medium (CM) was collected. The monocyte, macrophage, and LPS-induced macrophage CM were evaluated for IL-6 and tumor necrosis factor-alpha (TNF-α), and nitric oxide (NO) content using ELISA and the Griess Reagent System, respectively. Human primary (HN18, HN30, and HN4) and metastatic (HN17, HN31, and HN12) HNSCC cell lines were treated with the monocyte, macrophage, and LPS-induced macrophage CM. The proliferation and invasion of the HNSCC cell lines were evaluated using MTT and modified Boyden chamber assays, respectively.
Macrophages demonstrated increased IL-6 and CD14 expression. The P. gingivalis LPS significantly induced macrophage NO secretion, however, that of TNF-α decreased. The LPS-induced macrophages CM inhibited HN4 proliferation. Interestingly, the LPS-induced macrophage CM promoted invasion of all HNSCC cell lines.
Our data demonstrate that P. gingivalis LPS-induced macrophages increased NO secretion. The activated macrophage CM inhibited HN4 cell proliferation and promoted invasion of all HNSCC cell lines.
本研究旨在探讨牙龈卟啉单胞菌脂多糖（LPS）诱导的巨噬细胞对头颈部鳞状细胞癌（HNSCC）细胞系增殖和侵袭的影响。
用 12.5ng/ml 佛波醇 12-肉豆蔻酸 13-乙酸处理 THP-1 单核细胞 48 小时，将其向巨噬细胞分化。采用实时 PCR 和 ELISA 分别检测单核细胞和巨噬细胞白细胞介素 6（IL-6）mRNA 和细胞因子的表达。用免疫荧光标记法分析细胞 CD14 的表达。用 1μg/ml 牙龈卟啉单胞菌 LPS 诱导巨噬细胞 24 小时，收集条件培养基（CM）。用 ELISA 和格里塞试剂系统分别检测单核细胞、巨噬细胞和 LPS 诱导的巨噬细胞 CM 中 IL-6 和肿瘤坏死因子-α（TNF-α）和一氧化氮（NO）的含量。用人原发性（HN18、HN30 和 HN4）和转移性（HN17、HN31 和 HN12）HNSCC 细胞系处理单核细胞、巨噬细胞和 LPS 诱导的巨噬细胞 CM。用 MTT 和改良 Boyden 室测定法分别评估 HNSCC 细胞系的增殖和侵袭。
巨噬细胞表现出增加的 IL-6 和 CD14 表达。牙龈卟啉单胞菌 LPS 显著诱导巨噬细胞 NO 分泌，但 TNF-α减少。LPS 诱导的巨噬细胞 CM 抑制 HN4 增殖。有趣的是，LPS 诱导的巨噬细胞 CM 促进了所有 HNSCC 细胞系的侵袭。
我们的数据表明，牙龈卟啉单胞菌 LPS 诱导的巨噬细胞增加了 NO 的分泌。激活的巨噬细胞 CM 抑制了 HN4 细胞的增殖，并促进了所有 HNSCC 细胞系的侵袭。
