Isolation and identification of lung cancer stem like cells from human lung adenocarci-noma cell lines / 中国肿瘤临床

ABSTRACT

Objective:To isolate cell spheres containing cancer stem cells (CSCs) from lung cancer cell lines (A549 and H1299) and iden-tify their biological characteristics. Methods:By adopting the method of serum-free suspension culture, A549 cells and H1299 cells were cultured on no-adhesion plate to form tumor spheres. Clone assay, CCK-8 assay, and Transwell assay were employed to observe proliferation, self-renewal and invasion of tumor spheres. Besides, RT-PCR was performed to compare the expression levels of stem cell markers between sphere cells and adherent cells. Adherent A549 cells and A549 cell spheres were inoculated subcutaneously in nude mice and the tumor growth was assessed. Results:Isolated CSCs from A549 cells and H1299 cells in serum-free medium (SFM) without adhesion could grow as floating cell spheres. The results demonstrated that the self-renewal, proliferation and invasion of A549 and H1299 sphere cells were stronger than parent cells (P<0.05). When compared with adherent cells, the mRNA level of expres-sion of cell spheres' stem cell markers (Oct4 and Nanog) were significantly high (P<0.05) and A549 spheres had a stronger tumorigenici-ty in nude mice. Conclusion:SFM without adhesion can be a quick and easy method to construct stem cells model from human lung adenocarcinoma cell lines A549 and H1299.