| Objective: Ovarian cancer is the leading cause of death from gynec- ological cancers. The overall 5-year survival is therefore only 30%. The cytokine gene therapy is one of the biotherapeutic strategies for tumor, more and more attention has been paid in it. CD40, which from the tumor necrosis factor receptor superfamily, is present in variety of immune cells and tumor cells. Study has been demonstrated, CD40 ligand can cause direct activation of cell apoptosis or death in human colon cancer, breast cancer and lung cancer, which express CD40. This study is investigated the human soluble CD40 ligand (soluble CD40 ligand, sCD40L) in human epithelial ovarian cancer cell line SKOV3 and its cisplatin-resistant strains of SKOV3/DDP proliferation and possible mechanism of action, and to discussed the impact of drug resistance.Methods:1 RT-PCR was used to analyze the expression of CD40 gene in SKOV3 and SKOV3 /DDP cells.2 The effect of sCD40L and cisplatin on the proliferation of SKOV3 cells and SKOV3/DDP cells was measured by MTS colorimetric method.3 The effect of sCD40L combined with cisplatin on the proliferation of SKOV3 cells and SKOV3/DDP cells was measured by MTS colorimetric method.4 Cell cycles were assessed by flow cytometry.5 RT-PCR was used to analyze the expression of survivin mRNA, caspase-3 mRNA and caspase-9 mRNA in SKOV3 cells before and after sCD40L and sCD40L combined with cisplatin treatment.6 RT-PCR was used to analyze the expression of MDR1 mRNA, LRP mRNA, GST-πmRNA and survivin mRNA in SKOV3/DDP cells before and after sCD40L and sCD40L combined with cisplatin treatment.Results:1 CD40 mRNA were both expressed on SKOV3 and SKOV3/ DDP cells. The expression level of SKOV3 cell was 0.281±0.007, SKOV3/ DDP cell was 0.427±0.012.2 sCD40L inhibited the proliferation and induced apoptosis of ovarian cancer cells on dose depending manner, and the differences among different concentration were significant (P<0.05). The same concentration on two cell lines were not significant (P>0.05). The IC50 for sCD40L of SKOV3 and SKOV3/DDP were 3.47 and 3.22μg/ml.3 The IC50 for cisplatin of SKOV3 and SKOV3/DDP was 5.52 and 25.12μg/ml.The resistance multiple was 4.55.4 In the combined group, on SKOV3 cells, the IC50 of cisplatin was 5.52 to 1.46μg/ml, on SKOV3/DDP cells, the IC50 of cisplatin was 9.25μg/ml in the low dose group and 4.83μg/ml in the high dose sCD40L group. The reversion multiple was 2.71 and 5.20.5 After treated with sCD40L and cisplatin for 48h, the ratio of S phase of SKOV3 cell was decline, cell mainly arrested in G0/G1 phase by flow cyto- metry (P<0.05), and the percentage raised up, which were in dose-dependent manner. These were not manifest in cells with cisplatin 2.5μg/ml detected.6 SKOV3 cells were treated with different concentrations of drug inter- vention, with the drug concentration, the dose level of survivin mRNA expression was gradually decreased, caspase-3 mRNA and caspase-9 mRNA expression level were gradually increased. Compared with before treatment, the difference was statistically significant (P<0.05).7 SKOV3/DDP cells were treated with different concentrations of drug intervention, with the drug concentration, the dose level of MDR1 mRNA, LRP mRNA, GST-πmRNA and survivin mRNA expression were both gradually decreased. Compared with before treatment, the difference was statistically significant (P<0.05). Conclusions:1 CD40 were both expressed on SKOV3 and SKOV3/ DDP cells.2 sCD40L inhibits the proliferation of SKOV3 cells, which can enhance the inhibition after combined with cisplatin. sCD40L could decrease the gene expression of survivin in SKOV3 cells, and increase the gene expression of caspase-3 and caspase-9 to induce the apoptosis.3 sCD40L inhibits the proliferation of SKOV3/DDP cells.To some extent, the resistance can be reversed for SKOV3/DDP cells. sCD40L could decrease the gene expression of MDR1, LRP, GST-πand survivin in SKOV3/DDP. Therefore, the drug resistance of SKOV3/DDP to DDP would be reversed by sCD40L to some extend.
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