Spheroid formation induced by human lung adenocarcinoma cell line SPC-A1 and the tumorigenic ability of lung cancer stem cells

doi:10.3969/j.issn.2095-4344.2015.23.013

Abstract

Abstract:

BACKGROUND: There is no clear conclusion on whether the lung cancer stem cells can induce to spheroid formation and have tumorigenicity.
OBJECTIVE: To observe the spheroid formation induced by human lung adenocarcinoma cell line SPC-A1 and the tumorigenic ability of lung cancer stem cells.
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 cells. Immune fluorescence detection and PCR amplification were done to understand the expression of stem cell associated markers. NOD-SCID immunodeficient mice were subcutaneously implanted with lung spheroid cells to observe the tumor growth. In vivo fluorescence imager was used for radiography.
RESULTS AND CONCLUSION: After 5-10 days, lung spheroid cells were harvested. RT-PCR results showed that lung spheroid cells were positive for CD24, CD221, CCSP and SP-C. In addition, the lung spheroid cells and purified CD24⁺, CD221⁺lung cancer stem cells were both positive for TTF-1 of lung stem cells, OCT4 and Nanog of embryonic stem cells and TTF-1 of Bmi-1 lung stem cells. The fluorescence detection showed that over 80% lung spheroid cells expressed CCSP and OCT4; SPC-A1 cells had the characteristics of alveolar type II cells, and also expressed SP-C protein, but only about 5% of the cells expressed CCSP and OCT4. At 50 days after subcutaneous implantation of lung spheroid cells, in vivo fluorescence imaging showed that the diameter of tumor
in mice was 1 cm, indicating human lung adenocarcinoma cell line SPC-A1 can induce the spheroid formation, and lung cancer stem cells rich in the cell spheres have the tumorigenic ability.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Neoplastic Stem Cells, Lung Neoplasms

CLC Number:

R394.2

Hao Guang-jun, Liu Qing, Wang Juan, Ma Yan-e1 Ding Yan-hui. Spheroid formation induced by human lung adenocarcinoma cell line SPC-A1 and the tumorigenic ability of lung cancer stem cells [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(23): 3681-3685.

Figures/Tables 3

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