Spheroid formation induced by human lung adenocarcinoma cell line SPC-A1 and the tumorigenic ability of lung cancer stem cells
10.3969/j.issn.2095-4344.2015.23.013
- VernacularTitle:人肺腺癌细胞株SPC-A1诱导球体形成及肺癌干细胞致瘤能力评估
- Author:
Guangjun HAO
;
Qing LIU
;
Juan WANG
;
Yane MA
;
Yanhui DING
- Publication Type:Journal Article
- Keywords:
Stem Cells;
Neoplastic Stem Cells;
Lung Neoplasms
- From:
Chinese Journal of Tissue Engineering Research
2015;(23):3681-3685
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:There is no clear conclusion on whether the lung cancer stem cels can induce to spheroid formation and have tumorigenicity. OBJECTIVE: To observe the spheroid formation induced by human lung adenocarcinoma cel line SPC-A1 and the tumorigenic ability of lung cancer stem cels. METHODS:SPC-A1 at proliferating phase was cultured in serum-free DF12 culture medium, and then recombinant human insulin-like growth factor-1, recombinant human epidermal growth factor, and recombinant human fibroblast growth factor-10 were added to induce spheroid cels. Immune fluorescence detection and PCR amplification were done to understand the expression of stem cel associated markers. NOD-SCID immunodeficient mice were subcutaneously implanted with lung spheroid cels to observe the tumor growth.In vivo fluorescence imager was used for radiography. RESULTS AND CONCLUSION:After 5-10 days, lung spheroid cels were harvested. RT-PCR results showed that lung spheroid cels were positive for CD24, CD221, CCSP and SP-C. In addition, the lung spheroid cels and purified CD24+, CD221+ lung cancer stem cels were both positive for TTF-1 of lung stem cels, OCT4 and Nanog of embryonic stem cels and TTF-1 of Bmi-1 lung stem cels. The fluorescence detection showed that over 80% lung spheroid cels expressed CCSP and OCT4; SPC-A1 cels had the characteristics of alveolar type II cels, and also expressed SP-C protein, but only about 5% of the cels expressed CCSP and OCT4. At 50 days after subcutaneous implantation of lung spheroid cels, in vivo fluorescence imaging showed that the diameter of tumor in mice was 1 cm, indicating human lung adenocarcinoma cel line SPC-A1 can induce the spheroid formation, and lung cancer stem cels rich in the cel spheres have the tumorigenic ability.
