肿瘤相关巨噬细胞调控基质金属蛋白酶促口腔鳞癌细胞侵袭转移的实验研究

Effects of TAMs on oral squamous cancer cell invasion through enhancing production of MMPs

目的:探讨肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)对口腔鳞癌细胞基质金属蛋白酶(matrix metallproteinases,MMPs)表达以及其对口腔鳞癌细胞侵袭转移的影响。方法:用佛波脂(phorbol 12-myristate 13-acetate,PMA)和巨噬细胞集落刺激因子(macrophage colony-stimulating factor,M-CSF)刺激人单核/巨噬细胞株THP-1促其形成TAMs,收集其上清液作为TAMs条件培养基培养口腔鳞癌细胞,应用实时定量RT-PCR、Western blot方法检测TAMs条件培养基作用前后口腔鳞癌细胞MMPs表达的差异,应用Transwell侵袭实验检测口腔鳞癌细胞侵袭转移能力的差异。结果:THP-1细胞在PMA和MCSF作用后贴壁生长,分化成TAMs。口腔癌细胞在TAMs条件培养基作用48 h后,MMP-2、MMP-9、MMP-13 mRNA的表达水平显著增高,相同条件下蛋白质的表达水平也明显增高。TAMs条件培养基作用24 h后的口腔鳞癌细胞侵袭转移能力显著增强。结论:TAMs可以上调口腔鳞癌中MMPs的表达并促进其侵袭转移。

Objective：To explore the influence of tumor-associated macrophages （TAMs）on the expression of matrix metalloprotei-nases （MMPs）and the ability of invasion and metastasis of oral squamous cancer cells.Methods：We used phorbol 12-myristate 13-acetate （PMA）and macrophage colony-stimulating factor （M-CSF）to make THP-1 monocytes differentiate into TAMs,and then we col-lected the culture media of TAMs as condition medium （CM）to stimulate oral squamous cancer cells.Real-time RT-PCR and Western blot were applied to detect the expression of MMPs before and after stimulation of TAMs.The ability of invasion and metastasis of oral squamous cells was measured by Transwell assays.Results：When treated with PMA and M-CSF,THP-1 cells became attached,and differentiated into TAMs.Compared to control group,the expression of MMP-2,MMP-9,MMP-13 mRNA in experimental group were up-regulated when treated with TAMs CM for 48h.Protein level of MMP-2,MMP-9,MMP-13 was up-regulated accordingly.And stim-ulation of TAMs CM for 24h increased invasion of oral cancer cells.Conclusions：TAMs may promote the invasion and metastasis of o-ral squamous cancer cells through enhancing production of MMPs.