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目的探索人肺癌NCI-H446细胞系干细胞球体的培养,并鉴定其生物学特性。方法用无血清培养基培养人肺癌NCI-H446细胞得到肿瘤细胞球。将肿瘤细胞球传代扩增,并用含血清培养基培养促使其分化;四甲基偶氮唑盐(MTT)比色检测肿瘤球和普通NCI-H446细胞的增殖能力,并将肿瘤球和普通NCI-H446细胞分别植入裸鼠皮下,观察肿瘤形成;Transwell侵袭实验检测肿瘤球和普通NCI-H446细胞的侵袭能力的不同;流式细胞术检测CD133、CD44在肿瘤球和普通NCI-H446细胞中的表达,并筛选细胞表面标志物。结果在无血清培养基中,人肺癌NCI-H446细胞可以形成少量的肿瘤细胞球,并显示很强的自我更新和增殖能力,在含血清环境中能够诱导肿瘤球分化而贴壁生长;在动物实验中,接种5×105个细胞时,肿瘤球细胞较普通NCI-H446细胞显示更强的致瘤能力;在侵袭实验中,肿瘤球细胞的侵袭能力高于普通NCI-H446细胞;干细胞标志物CD133及CD44在肿瘤球细胞的表达较普通NCI-H446细胞明显增高。结论人肺癌细胞NCI-H446中存在癌干细胞,且可以通过无血清培养、分离和富集。
Objective To explore the culture of human lung cancer cell line NCI-H446 and to identify its biological characteristics. Methods Human lung cancer NCI-H446 cells were cultured in serum-free medium to obtain tumor cell spheres. The tumor cells were passaged and subcultured, and then differentiated by culture in serum-containing medium. The proliferation of tumor cells and normal NCI-H446 cells was detected by MTT colorimetric assay. H446 cells were implanted into the nude mice subcutaneously to observe the tumor formation. Transwell invasion assay was used to detect the invasion ability of tumor cells and normal NCI-H446 cells. CD133 and CD44 were detected by flow cytometry in tumor spheres and normal NCI-H446 cells And screen for cell surface markers. Results In the serum-free medium, human lung cancer NCI-H446 cells formed a small number of tumor cell spheres and showed strong self-renewal and proliferative ability, which induced tumor spheroid differentiation and adherent growth in a serum-containing environment. In experiment, tumor cells showed more tumorigenic ability than normal NCI-H446 cells when inoculated with 5 × 105 cells. Invasion ability of tumor cells was higher than that of normal NCI-H446 cells. Stem cell markers The expression of CD133 and CD44 in tumor cells was significantly higher than that in normal NCI-H446 cells. CONCLUSIONS: Cancer stem cells exist in human lung cancer cell line NCI-H446 and can be cultured, isolated and enriched by serum-free culture.