Hao G.-J.,Yulin Municipal First Hospital |
Liu Q.,Yulin Municipal First Hospital |
Wang J.,First Affiliated Hospital Of Xian Jiaotong University |
Ma Y.-E.,Yulin Municipal First Hospital |
Ding Y.-H.,Yulin Municipal First Hospital
Chinese Journal of Tissue Engineering Research | Year: 2015
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. © 2015, Journal of Clinical Rehabilitative Tissue Engineering Research, All Right Reserved.