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目的:在建立舌癌Tca8113淋巴道转移裸鼠模型的基础上,建立永生化的舌癌细胞系,并进一步探讨舌癌转移淋巴结组织中同源癌细胞相关标记物的表达情况,为舌癌干细胞研究提供基本的研究模型。方法:采用癌细胞足垫注射法建立淋巴道转移模型,4周后取淋巴结进行原代细胞培养并分离、纯化舌癌细胞进行连续传代培养、进行永生化细胞系的生物学特性鉴定。结果:在淋巴道转移模型中,HE染色与EMA免疫组化染色发现淋巴结有散在或灶性鳞状上皮癌细胞转移。进而取转移模型裸鼠的淋巴结组织进行原代细胞培养,经分离和纯化以及连续传代而建立舌癌永生化细胞系,命名为Tca8113-Ml,目前传代100余代,时间近2年,生长稳定,倍增时间为22.51h,染色体众数58,保持人染色体形态,细胞形态与Tca8113细胞相似。Tca8113-Ml在裸鼠体内的成瘤率为100%,裸鼠腋窝皮下成瘤的组织切片显示为典型的鳞状上皮细胞癌。结论:在舌癌淋巴道转移模型中,可分离培养与建立永生化的舌癌细胞系。
OBJECTIVE: To establish an immortalized tongue cancer cell line based on the establishment of tongue cancer Tca8113 lymph node metastasis nude mouse model and to further explore the expression of related markers of homologous cancer cells in tongue cancer metastatic lymph node tissue, Research provides the basic research model. Methods: Lymphatic metastasis model was established by footpad injection of cancer cells. After 4 weeks, lymph node was taken for primary cell culture and isolated. Tongue cancer cells were purified and subcultured. The biological characteristics of immortalized cell lines were identified. Results: In lymphatic metastasis model, HE staining and immunohistochemical staining with EMA revealed scattered or focal squamous cell carcinoma lymph node metastasis. Then the lymph node tissues of the metastatic nude mice were used for primary cell culture, and the immortalized tongue cancer cell line was established by isolation, purification and continuous passage and named as Tca8113-Ml. Currently, it has been passaged for more than 100 generations for nearly 2 years and has stable growth , Doubling time was 22.51h, chromosome number 58, to maintain human chromosome morphology, cell morphology and Tca8113 cells similar. The tumorigenic rate of Tca8113-Ml in nude mice was 100%. Tissue sections of subcutaneous tumor in axilla of nude mice showed typical squamous cell carcinoma. CONCLUSION: Immortalized tongue cancer cell lines can be isolated and cultured in lymphatic metastasis model of tongue cancer.