Isolation and identification of cancer stem-like cells from lung cancer H1299 cells

赵文思; 罗弋; 黎博一; 周寒静; 张涛

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Isolation and identification of cancer stem-like cells from lung cancer H1299 cells

更新时间：2026-01-09

- Isolation and identification of cancer stem-like cells from lung cancer H1299 cells
- Journal of Army Medical University Vol. 38, Issue 7, Pages: 708-713(2016)
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  CLC： R329-33;R730.23;R734.2
- Published：2016
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赵文思,罗弋,黎博一,周寒静,张涛.人肺癌H1299肿瘤干细胞样细胞的分离培养及鉴定[J].陆军军医大学学报(原第三军医大学学报),2016,38(07):708-713.

　Zhao Wensi,Luo Yi,Li Boyi,Zhou Hanjing,Zhang Tao.Isolation and identification of cancer stem-like cells from lung cancer H1299 cells[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(07):708-713.
赵文思,罗弋,黎博一,周寒静,张涛.人肺癌H1299肿瘤干细胞样细胞的分离培养及鉴定[J].陆军军医大学学报(原第三军医大学学报),2016,38(07):708-713. DOI：

　Zhao Wensi,Luo Yi,Li Boyi,Zhou Hanjing,Zhang Tao.Isolation and identification of cancer stem-like cells from lung cancer H1299 cells[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(07):708-713. DOI：

摘要

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（P0.05）。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。

Abstract

Objective To isolate and culture cancer stem-like cells (CSCs) from human lung cancer cell line H1299

and then identify their stem cell characteristics. Methods H1299 cells were cultured in serum-free medium under non-adherent conditions to induce the formation of tumorspheres. Immunofluorescent assay

flow cytometery

PCR and Western blotting were performed to evaluate the expression of CSCs markers

CD133 and ABCG2. The clonality and tumorigenicity in vivo were also examined. Results The tumorspheres were formed by sphere-forming culturing. The data demonstrated that there were stronger capacities of proliferation and tumorigencity in the tumorspheres derived from H1299 cells than the adherent cells. And the expression levels of CD133 and ABCG2 were significantly higher in the tumorspheres (P0.05). Conclusion The tumorspheres derived from H1299 cells by floating spheres culturing in serum-free condition are of high expression of CD133 and ABCG2

and possess stem cell characteristics.

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