Epithelial ovarian cancer cells: SKOV3 and its cisplatin-resisitant strain SKOV3/DDP cells were used to test the effect of sCD40L and cisplatin. The proliferation of SKOV3 and SKOV3/DDP cells were measured by MTT. Cell cycle was assessed by flow cytometry. The mRNA expressions of targeted genes were detected by qRT-PCR. The protein expressions were detected by Western blotting.
sCD40L showed a significant dose-dependence inhibitory effect on the proliferation of ovarian cancer cell lines. sCD40L in combination with cisplatin could sensitized SKOV3/DDP cells to cisplatin treatment and reversed the drug resistance of SKOV3/DDP cells. The reversal ratios of 1 μg/ml sCD40L combined with cisplatin in SKOV3 and SKOV3/DDP cells were 2.11, 2.71, while the reversal ratios of 2 μg/ml sCD40L combined with cisplatin in SKOV3 and SKOV3/DDP cells were 3.78, 5.20, respectively. sCD40L or sCD40L combined cisplatin increased tumor cells in G0/G1 phase. sCD40L in combination with cisplatin decreased the expression levels of GST-π, LRP, Survivin, p53 and Bcl-2 in both epithelial ovarian cancer cell lines. The protein expression level of GST-π, LRP and P53 protein was also decreased upon sCD40L in combination with cisplatin although the expression level of Bcl-2 and survivin protein had no significant difference.
sCD40L inhibits the proliferation of SKOV3 and SKOV3/DDP cells. The combined application of sCD40L and cisplatin can strength the inhibitory effect of cisplatin, and to a certain extent, reversing the resistance to cisplatin in SKOV3/DDP cells. sCD40L could lead a cell block in G0/G1 phase and make the cell growth restrained. sCD40L could induce SKOV3 and SKOV3/DDP cells apoptosis and reverse drug resistance through cutting GST-π mRNA, LRP mRNA, survivin mRNA, p53 mRNA and Bcl-2 mRNA and decreasing the expression of GST-π, LRP and P53 protein in SKOV3 and SKOV3/DDP cells, which provides in-vivo experiment basis to the application of sCD40L as a drug improving ovarian cancer cells sensitivity to cisplatin.