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目的通过原代培养人喉癌实体瘤细胞,克隆分离异质性细胞亚群,初步研究喉癌细胞肿瘤异质性。方法原代培养人喉癌细胞,应用有限稀释法对体外培养的喉癌细胞进行单细胞克隆分离异质性亚群,比较其在形态、分化能力、细胞周期、增殖能力及裸鼠异体移植成瘤能力等方面的差异。结果分离到1型和2型细胞形态差异最大的两个亚群。1型细胞成长梭形,突起少,亚克隆培养未产生子代细胞,裸鼠异体移植不成瘤(0/5);2型细胞成多角形,多突起,继续培养产生的子代克隆出现异质性细胞,裸鼠异体移植可成瘤(5/5),体外增值能力强,处于细胞周期G0/G1的比例明显高于1型细胞。结论原发性人喉癌实体瘤细胞具有异质性,2型细胞具有肿瘤干细胞的特性。
Objective To study the primary tumor cell heterogeneity of human laryngeal cancer cells by cloning and isolating heterologous cell subpopulations. Methods Primary human laryngeal carcinoma cells were cultured in vitro. Tissue cultures of laryngeal carcinoma cells were subjected to single cell cloning and isolation using a limiting dilution method. The morphology, differentiation ability, cell cycle, proliferation and nude mice allograft were compared. Differences in tumor abilities etc. As a result, two subpopulations with the largest differences in the morphology of type 1 and type 2 cells were isolated. Type 1 cells grow spindle-shaped with few projections. Subcloning cultures do not produce progeny cells. Allograft transplantation in nude mice does not become tumorous (0/5). Type 2 cells become polygonal and multi-projections. Different generations of clones resulting from continuous culture appear differently. In allogeneic nude mice, allogeneic mice can be tumorigenic (5/5), have a strong in vitro proliferation ability, and have a significantly higher ratio of cell cycle G0/G1 than type 1 cells. Conclusion The primary solid tumor cells of human laryngeal carcinoma are heterogeneous, and type 2 cells have the characteristics of cancer stem cells.