裸鼠体内原位移植法筛选粘液表皮样癌高转移细胞

目的：筛选高转移人涎腺粘液表皮样癌细胞。方法：用人涎腺粘液表皮样癌细胞Mc3经裸鼠涎腺移植、鼠间原位移植、细胞培养技术获取所需细胞，用相差显微镜、电子显微镜、细胞计数法、染色体显示法、流式细胞术、软琼脂克隆形成法以及癌细胞裸鼠尾静脉注射法研究其生物学特性。结果：经4次鼠间原位移植，肺转移率为3／10。取转移灶行细胞培养，获得形态均一、生长旺盛的上皮样细胞，命名为M3SP4。M3SP4细胞所致肺肿瘤显示粘液表皮样癌组织学特征，细胞染色体为亚二倍体，仍保持人类染色体形态。M3SP4与Mc3形态相似，群体倍增时间（h）分别为23.9和25.9，S期细胞（％）分别占细胞周期26.8和15.3，克隆形成率（％）分别为54.6和30.2，经尾静脉注射后M3SP4细胞引起的肺转移结节比Mc3多35％。结论：M3SP4是转移力强于Mc3的人涎腺粘液表皮样癌细胞系，Mc3细胞经裸鼠涎腺移植、鼠间原位移植后转移力有所提高。

Selection of highly metastatic cells of human salivary gland mucoepidermoid carcinoma by in situ transplantation of Mc3 cells in nude mice

Objective: To select highly metastatic cells from human salivary gland mucoepidermoid carcinoma cell clone Mc3. Methods: In situ transplantation of Mc3 cells into submandibular gland of nude mice, in situ transplantation of Mc3 induced lung metastasized tumor tissue among nude mice and cell culture were employed to obtain the wanted cells. Morphological observation, cell growth analysis, flow cytometry, chromosome staining, clonogenic assay and artificial metastasis test in nude mice were used to characterize the cells. Results: Lung metastasis was observed in 3 out of 10 nude mice after 4 cycles of in situ transplantation of Mc3 cell induced lung metastasized tumor tissue. Epidermoid cells with similar morphology to Mc3 were obtained through cell culture and the cells were named M3SP4. M3SP4 cell induced lung metastatic foci were histologicaly proved to be mucoepidermoid carcinoma. Subdiploid karyotype with human chromosome morphology was observed in M3SP4 and Mc3 cells. The population doubling time (h) of M3SP4 and Mc3 cells was 23.9 and 25.9, the percentage of S phase cells in cell cycle 26.8 and 15.3, clonogenecity (%) 54.6 and 30.2, respectively. The artificial lung metastatic potential of M3SP4 cells was 35% higher than that of Mc3 in nude mice. Conclusion: M3SP4 cells are of human mucoepidermoid carcinoma with higher metastatic potential than Mc3. In situ transplantation of mucoepidermoid carcinoma cells or lung metastasized tumor tissue may maintain the metastatic potential of the cells.