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目的:探讨神经酰胺增强卵巢癌顺铂敏感性的作用及机制,为筛选有效的卵巢癌顺铂耐药逆转剂提供实验依据。方法:本研究采用MTT法测定不同剂量的C6-神经酰胺(Ceramide C6)(5、10、20、40μmol/L)对卵巢癌顺铂化疗的增敏作用,以RT-PCR、Westernblot法检测Ceramide C6对肿瘤细胞Bcl-2 mRNA及其蛋白表达的影响,应用流式细胞技术观察Ceramide C6对肿瘤细胞凋亡的影响。结果:Ceramide C6对SKOV3、SKOV3/DDP细胞生长有抑制作用,与DDP联合应用时可增强DDP敏感性。SKOV3细胞中,与对照组相比,不同浓度Ceramide C6使顺铂敏感性分别增加了7.23%、20.14%、45.02%、57.75%。SKOV3/DDP细胞中,顺铂敏感性分别增加了23.08%、24.57%、45.86%、65.78%。随Ceramide C6浓度增加,SKOV3和SKOV3/DDP细胞中Bcl-2 mRNA及蛋白表达逐渐减低,差异有统计学意义(F=41.780,P=0.000;F=223.258,P=0.000;F=98.306,P=0.000;F=224.687,P=0.000)。Ceramide C6可增加SKOV3、SKOV3/DDP细胞凋亡率,差异有统计学意义(F=135.625,P=0.000;F=224.906,P=0.000)。结论:神经酰胺可能通过抑制肿瘤细胞Bcl-2 mRNA及其蛋白表达,诱导肿瘤细胞凋亡途径,增强顺铂对卵巢癌SKOV3和SKOV3/DDP细胞体外生长的抑制作用,在一定程度上逆转了卵巢癌顺铂耐药。
OBJECTIVE: To investigate the effect and mechanism of ceramide on cisplatin sensitivity in ovarian cancer and to provide experimental evidence for screening effective cisplatin-resistant reversal agents in ovarian cancer. Methods: MTT assay was used to determine the sensitizing effect of different doses of Ceramide C6 (5, 10, 20, 40μmol / L) to cisplatin chemotherapy in ovarian cancer. RT-PCR and Western blotting were used to detect Ceramide C6 on the expression of Bcl-2 mRNA and protein in tumor cells. The effect of Ceramide C6 on the apoptosis of tumor cells was observed by flow cytometry. Results: Ceramide C6 inhibited the cell growth of SKOV3 and SKOV3 / DDP cells and increased the sensitivity of DDP when used in combination with DDP. In SKOV3 cells, Ceramide C6 increased cisplatin sensitivity by 7.23%, 20.14%, 45.02% and 57.75%, respectively, compared with the control group. The sensitivity of cisplatin to SKOV3 / DDP cells increased by 23.08%, 24.57%, 45.86% and 65.78% respectively. With the increase of Ceramide C6 concentration, the expression of Bcl-2 mRNA and protein in SKOV3 and SKOV3 / DDP cells decreased gradually (F = 41.780, P = 0.000; F = 223.258, P = 0.000; F = 98.306, P = 0.000; F = 224.687, P = 0.000). Ceramide C6 increased the apoptosis rate of SKOV3 and SKOV3 / DDP cells, the difference was statistically significant (F = 135.625, P = 0.000; F = 224.906, P = 0.000). CONCLUSION: Ceramide may reverse the ovary to a certain degree by inhibiting the expression of Bcl-2 mRNA and its protein, inducing apoptosis of tumor cells and increasing the inhibitory effect of cisplatin on the growth of ovarian cancer SKOV3 and SKOV3 / DDP cells in vitro Cancer cisplatin resistance.