中国美容医学2013，Vol.22Issue(15)：1607-1611,5.
内皮前体细胞与口腔鳞癌细胞自发性融合的实验研究
Spontaneous fusion of endothelial progenitor cells and oral squamous cancer cells
孙毅 1尚政军1
作者信息
- 1. 武汉大学口腔医院第一颌面外科 湖北武汉 441003
- 折叠
摘要
Abstract
Objective To investigate whether other bone marrow derived cells besides BMSCs can spontaneous fuse with oral squamous cancer cells.Methods Mononuclear cells (MNCs) were obtained from 5ml fresh human peripheral blood by density gradient fractionation,and resuspended in complete medium (EGM-2 MV) supplemented with 10％ FBS and seeded on 24-well culture plates.PKH26 labeled endothelial progenitor cells were cocultured with CFSE labeled oral squamous cancer cell lines Cal 27,SCC 4 and SCC 9.After 24 h post-coculture,bi-positive cells including two or more nucleus were identified as hybrid cells.Flow cytometry was used to analyze the fusion efficiency of three oral squamous cancer cell lines with EPCs.Immunofluorescence Staining was used to detect the epithelial molecule Cytokeratin and mesenchymal marker Vimentin.Results Single-cell clone appeared after 8～12 days were endothelial-like in appearance and formed a cobblestone-shaped cell monolayer.After 24 h post-coculture,PKH26/CFSE bi-fluorescent cells contained two or more nucleus can be observed in the coculture system.Flow cytometry analysis showed that the fusion efficiency of either oral squamous cancer line with EPCs was 0.62 ±0.028,0.59 ±0.057,0.65 ±0.099,respectively and no significant different was observed between the oral squamous cancer cell lines used (P=0.706).Immunofluorescence Staining revealed that Cytokeratin/Vimentin bi-positive cells contained two different nuclei were existed in the coculture system.Conclusion EPCs can spontaneously fuse with oral squamous cancer cells and the hybrid cells coexpressed the Cytokeratin and Vimentin.The present study provided a novel strategy to understand the EMT and tumor invasion.关键词
内皮前体细胞/细胞融合/口腔鳞癌细胞/PKH26/CFSEKey words
endothelial progenitor cells/cell fusion/oral squamous cancer cells/PKH26/CFSE分类
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孙毅,尚政军..内皮前体细胞与口腔鳞癌细胞自发性融合的实验研究[J].中国美容医学,2013,22(15):1607-1611,5.
