The biological characteristics of holoclone from ovarian cancer cell SKOV3 and its ability to form vasculogenic mimicry / 肿瘤

ABSTRACT

Objective: To observe the biological characteristics of holoclone cells which were sperated by single cell clone from hunman epithelial ovarian cancer SKOV3 cells and its ability to from vasculogenic mimicry (VM) formation. Methods: SKOV3 cells were cultured and separated into three different single cell clones by limited dilution method. The tumor formation test by in nude mice in vivo and soft agar colony formation test were used to compare the biological characteristics of the three kinds of single cell clones. Three-dimensional cell culture model was established to observe the ability of VM formation in three separated cell clones. The expression levels of CD1 33, matrix metalloproteinases-2 (MMP-2) and MMP-9 in holoclone cells, meroclone cells, paraclone cells and parental cells were detected by Western blotting. Results: Single cell clones were cultured for 10-14 days, and the average proportions of the three cell clones were (17.4±0.8) % (holoclone), (30.1 ±0.4) % (meroclone), and (52.5±1.1) % (paraclone), respectively. After three different cell clones were subcutaneously inoculated in female nude mice, the rate of tumor formation of holoclone was 80%, but those of meroclone and paraclone were both 0. The average cell clone formation rates of three cell clones were (16.7 ± 3.8) %, (6.5±1.5) % and (1.2±0.7) %, respectively (P < 0.05). Under the condition of three-dimensional cell cultivation, holoclone cells could form VM, whereas meroclone and paraclone cells couldn't. The expression levels of CD133, MMP-2 and MMP-9 proteins in holoclone group were significantly higher than those of the maternal SKOV3 cells group, and also higher than those of the meroclone and paraclone groups (P < 0.05). Conclusion: Three kinds of single cell clones have heterogeneousness in morphology and biological characteristics, and holoclone cells have the characteristics of tumor stem-like cells, which can form VM. Holoclone may be a SKOV3 cell subset with the higher ability of recurrence.