小鼠口腔癌淋巴道转移模型骨髓播散肿瘤细胞的检测

目的：检测小鼠口腔癌淋巴道转移模型癌变过程不同时期的骨髓播散肿瘤细胞（disseminated tumor cell,DTC）,探讨小鼠口腔癌变过程与骨髓DTC的关系.方法：病理检查明确诊断为正常舌黏膜、舌单纯性增生的小鼠各10只,舌轻中度异常增生、舌重度异常增生、舌鳞癌的小鼠各20只.采用Ficoll密度梯度离心法提取小鼠股骨骨髓里单个核细胞并制成细胞涂片,免疫细胞化学（Immunocytochemistry,ICC）检测骨髓DTC,比较其数目差异.结果：舌重度异常增生8只、舌鳞癌组13只发现DTC,阳性率分别为40％（8/20）和65 ％（13/20）,每100个单核细胞中DTC数分别为3.03±0.75个和5.20±0.74个,差异均有统计学意义（P＜0.05）.其余各组均未发现DTC（0/10）.结论：小鼠舌黏膜恶变过程中,舌重度异常增生时已出现DTC,并随病变程度加重,骨髓DTC的发生率及其细胞数量增加.

Detection of disseminated tumor cells in bone marrow of mouse oral carcinoma model

Objective： To detect disseminated tumor cells （DTC） in bone marrow of mouse oral carcinoma model and investigate the relationship between different stages of carcinogenesis of the oral mucosa and DTC. Method： The femur bone marrows, which were selected from 10 animals with nomal tongue mucosa and 10 with simple hyperplasia tongue mucosa, 20 with mild-to-moderate dysplasia, 20 with severe abnormal hyperplasia and 20 with squamous cell carcinoma, were collected. Single nuclear cells were extracted by Ficoll density gradient centrifugation, and DTC were detected by Immunocytochem- istry with epithelial-specific cytokeratin antibodies. Result： Eight cytokeratin positive （CK＋） cells were found in 20 （40 %） cases of severe dysplasia and 13 in 20 （65 %） cases of squamous cell carcinoma. The mean number of CK＋ cells per 100 single nuclear cells in severe dysplasia and squamous cell carcinoma were 3.03±0.75 and 5.20 ±0.74, respectively. There was significant difference between two groups （P 〈0.05）. However, CK＋ cells were not found in the rest groups （0 / 10）. Conclusion：There may be DTC in the stage of tongue severe dysplasia, and the rate and number of DTC increase from the stage of severe dysplasia on up to the stage of squamous cell carcinoma.