Liu Y, Hu T, Shen J, Li S F, Lin J W, Zheng X H, Gao Q H, Zhou H M
Department of Oral Medicine, West China College of Stomatology, Sichuan University, Chengdu, China.
Oral Dis. 2006 Jul;12(4):375-80. doi: 10.1111/j.1601-0825.2005.01207.x.
Carcinoma-associated fibroblasts (CAFs) have been suggested to regulate the initiation and progression of many types of solid tumors. The aim of the study was to separate, cultivate, identify oral CAFs, and to investigate their biological characteristics.
The primary CAFs and normal fibroblasts (NFs) of the tongue were obtained by tissue culture. Then cells were dissociated by 0.25% trypsin and purified by curettage method combining with trypsinization. The cells were verified according to morphological observation and immunohistochemical staining of certain proteins. Multiple proliferation indexes and karyotype of the cells were assayed.
Third passage purified oral CAFs and NFs were attained successfully. The morphological characteristics of the CAFs changed significantly comparing to the NFs. The CAFs showed positive staining for vimentin, alpha-smooth muscle actin and matrix metalloproteinases-2. The proliferation and mitosis ability of the CAFs were significantly increased compared with the NFs (P < 0.05). No karyotypic abnormalities were found in the CAFs.
There were obvious differences in the biological characteristics between oral CAFs and NFs. The results may provide us an experimental foundation for further studies on the roles of CAFs in the initiation and progression of oral cancer.
癌相关成纤维细胞（CAFs）被认为可调节多种实体瘤的发生和发展。本研究旨在分离、培养、鉴定口腔CAFs，并研究其生物学特性。
通过组织培养获取舌部原代CAFs和正常成纤维细胞（NFs）。然后用0.25%胰蛋白酶消化细胞，采用刮除法结合胰蛋白酶消化法进行纯化。根据形态学观察和某些蛋白质的免疫组织化学染色对细胞进行鉴定。检测细胞的多种增殖指标和核型。
成功获得第三代纯化的口腔CAFs和NFs。与NFs相比，CAFs的形态特征发生了显著变化。CAFs波形蛋白、α-平滑肌肌动蛋白和基质金属蛋白酶-2染色呈阳性。与NFs相比，CAFs的增殖和有丝分裂能力显著增强（P<0.05）。CAFs未发现核型异常。
口腔CAFs和NFs的生物学特性存在明显差异。该结果可为进一步研究CAFs在口腔癌发生和发展中的作用提供实验基础。
