Influence of cocl2 in cisplatin sensitivity of human ovarian cancer skov3 cells / 吉林大学学报(医学版)

ABSTRACT

Objective: To observe the effect of CoCl on the cisplatin sensitivity of human ovarian cancer SKOV3 cells, and to clarify the possible mechanism. Methods: The SKOV3 cells were Cultured in vitro and randomly divided into control group, CoCT group, cisplatin (DDP) group and CoCT combined with DDP (combination) group. The cells in CoCL group were Cultured in normal cell medium for 20 h after cultured in 200 pmol • L CoCL for 4 h, the cells in DDP group were cultured in normal cell medium containing 10 mg • L DDP for 24 h, and the cells in combination group were cultured in 10 mg • L DDP for 20 h after cultured in 200 //mol • L CoCl • for 4 h. The survival rates of SKOV3 cells in various groups were detected by MTT method, and the positive expression intensities of hypoxia-inducible factor-1 a ( HIF-la) and inducible nitric oxide synthase (iNOS) in the cells in various groups were detected by immunofluorescence method. Rhod 2-AM fluorescence probe was used to observe the levels of Ca in mitochondria in the cells in various groups. Western blotting method was used to observe the expression levels of cytochrome C (cyto C) cysteinyl aspartasc 3 (caspasc 3) and cleaved cysteinyl aspartasc 3 (cleaved caspase 3). Muse apoptosis assay kit was used to detect the apoptotic rates of cells in various groups. Results: Compared with control group, the survival rate of the cells in CoCI group had no significant change (P> 0.05). and the survival rates of the cells in DDP and combination groups were decreased ( P0. 05) . and the expression levels of cyto C. caspase 3 and cleaved caspase 3 in DDP group were increased significantly ( P < 0.05); comparexl with DDP group, they were lower than those in combination group ( P<0. 05). Comparexl with control group∗ the apoptotic rate of SKOV3 cells in DDP group was increased significantly (P<.0. 05); the apoptotic rate of SKOV3 cells in combination group was lowe'r than that in DDP group (P<0. 05). Conclusion: CoCI can redece the mitochondrial apoptosis of human ovarian cancer SKOV3 cells by inhibiting the DDP-inducexl enhancement of iNOS expression and dccrease the sensitivity of SKOV3 cells to cisplatin.