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目的从人肺癌H1299细胞系中分离培养肺癌干细胞样细胞,并进一步鉴定其干细胞特性。方法在低黏附条件下无血清诱导培养H1299细胞,富集H1299肿瘤细胞球,利用免疫荧光、流式细胞仪、PCR及Western blot检测干性标志物CD133和ABCG2的表达,并观察克隆形成和体内致瘤能力的改变。结果肺癌H1299细胞经无血清诱导培养可形成悬浮的细胞球。与普通贴壁H1299细胞相比,细胞球具有更强的增殖能力和致瘤性,且高表达干性基因CD133和ABCG2,差异有统计学意义(P<0.05)。结论运用无血清悬浮细胞球培养方法富集的肺癌H1299细胞系肿瘤球高表达CD133和ABCG2,具有干细胞特性。
Objective To isolate and culture lung cancer stem cell-like cells from human lung cancer cell line H1299 and further identify their stem cell characteristics. Methods H1299 cells were cultured in serum - free medium with low adhesion and H1299 tumor cells were enriched. The expression of CD133 and ABCG2 were detected by immunofluorescence, flow cytometry, PCR and Western blotting. Clonal formation and in vivo Tumor growth ability changes. Results Lung cancer H1299 cells were cultured in serum-free medium to form suspended cells. Compared with the normal adherent H1299 cells, the cytoplasm had stronger proliferation and tumorigenicity, and the expression of CD133 and ABCG2 was higher than that of the normal adherent H1299 cells (P <0.05). Conclusion The tumor spheres of H1299 cell line enriched in serum-free suspension cell culture method express CD133 and ABCG2 highly and have stem cell characteristics.