Identification of biological characteristics of human ovarian cancer cell line HO8910-derived stem cells

FANG Li-sha; XU Jun; QIU Ying-ying; YAN Hong-chao; LIN Xiao-man; HUANG Hong-xiang; HAN Qiu-yu

doi:10.3969/j.issn.1002-266X.2015.36.006

Abstract：Objective To identify the biological characteristics of human ovarian cancer cell line HO8910-derived stem cells.Methods The pre-screening of ovarian cancer cell line HO8910-derived stem cells were subcultured ( HO8910 cells were used as the control group) in serum-free medium.The capacities of forming spheroids and self-renew were ob-served.Then ovarian cancer stem cells ( CSCs) were seeded in medium containing serum and cultured to observe the differ-entiation capacity, and the stem cell-specific markers were also tested.We tested the sensitivity of stem cells to cisplatin, doxorubicin and mitoxantrone by using drug susceptibility test.Finally, we inoculated the ovarian CSCs after passaging from culturing in serum-free media to NOD/SCID ( non-obese diabetic/severe combined immunodeficient mice) mice in order to observe the tumorigenicity in vivo.Results The forming spheroid capacity of HO8910 cells in the serum-free medium was significantly lower than that of the ovarian cancer cells (P<0.05).The positive expression rates of CD133 and CD117 in the ovarian CSCs before differentiation were higher than those after differentiation (all P<0.05).The expression of AB-CG2, Nanog, Oct4, BCRP and E-cadherin in the ovarian CSCs after differentiation was lower than that before differentia-tion (all P<0.05).No statistical significance was found in the above indexes before and after HO8910 cells differentiation (all P>0.05).The relative activities in the ovarian CSCs were higher than in HO8910 cells when they were respectively treated with 0.25 and 0.5μg/mL cisplatin, 0.5 and 1.5μmol/L doxorubicin and 0.05 and 0.25μg/mL mitoxantrone for 48 h (all P<0.05).The tumor formed when the nude mouse was inoculated with 1 ×103 ovarian CSCs.With the increase of inoculated stem cells, the tumor formation ratio was gradually increased ( all P<0.05 ) , and the time was gradually shortened (all P<0.05).Conclusion The ovarian CSCs derived from human ovarian cancer cell line HO8910 have the abilities of self-renew, differentiation, in vivo tumorigenicity, highly expressed stem cell genes and multidrug resistance, which meet the standard of tumor stem cell theory.