褪黑素联合顺铂对卵巢癌SKOV3细胞的生长抑制作用

目的：探讨褪黑素（MLT）、顺铂（DDP）单独及联合应用对人卵巢癌SKOV3细胞的生长抑制作用，为MLT作为卵巢癌化疗辅助药物的研究提供理论依据。方法：将体外培养至对数生长期的卵巢癌SKOV3细胞分为对照组、单用MLT组（0.5、1.0和2.5 mmol/L）、单用DDP组（25和50 mg/L）、联合应用MLT及DDP组（MLT 0.5 mmol/L联合DDP 25 mg/L； MLT 0.5 mmol/L联合DDP 50 mg/L；MLT 1.0 mmol/L联合DDP 25 mg/L；MLT 1.0 mmol /L联合DDP 50 mg/L；MLT 2.5 mmol/L联合DDP 25 mg/L；MLT2.5 mmol/L联合DDP 50 mg/L）。应用显微镜观察法、MTT法测定用药后卵巢癌SKOV3细胞的凋亡及增殖抑制率，并采用免疫化学方法测定卵巢癌SKOV3细胞用药后的血管内皮生长因子（VEGF）的表达。结果：显微镜下观察结果显示用药组细胞贴壁能力减弱、变小变圆、折光性减弱、细胞皱缩、核固缩、染色质凝集、核碎裂。联合用药组凋亡细胞增多；MTT 法检测结果显示0.5、1.0和2.5 mmol/L-1的MLT单独应用或与25和50 mg/L的DDP联合应用均有抑制细胞生长作用（P<0.05），且随浓度升高抑制率升高，联合用药组两药物间均有协同作用，两药相互作用指数（CDI）＜1；免疫化学染色法显示单用药组及联合用药组VEGF表达较对照组均降低，联合用药组VEGF表达明显降低（P<0.05）。结论：MLT对卵巢癌细胞生长有抑制作用，MLT联合DDP对VEGF表达下调有协同作用，提示适当浓度的MLT配伍DDP作用于卵巢癌细胞SKOV3可达到较好疗效。

Growth inhibiting effect of melatonin combined with cisplatin on ovarian cancer cell line SKOV3

Objective To study the inhibitory effects of melatonin (MLT),cisplatin (DDP),MLT combined with DDP on human ovarian cancer cell line SKOV3,and provide the theoretical basis for the study on MLT as an ancillary drug of ovarian cancer.Methods The ovarian cancer cell SKOV3at logarithmic growth phase were divided into control group,MLT groups(0.5,1.0and 2.5mmol·L-1),DDP groups(25,50mg·L-1)and MLT combined with DDP groups (MLT 0.5mmol·L-1 combined with DDP 25mg·L-1;MLT 0.5mmol·L-1 combined with DDP 50mg·L-1;MLT 1.0mmol·L-1 combined with DDP 25mg·L-1;MLT 1.0mmol·L-1 combined with DDP 50mg·L-1;MLT 2.5mmol·L-1 combined with DDP 25mg·L-1;MLT 2.5mmol·L-1 combined with DDP 50mg·L-1).The apoptotic rate and proliferation inhibitory rate of ovarian cells were detected with microscopic observation and MTT assay,and the expression of VEGF in ovarian cancer cell SKOV3was determined.Results Observing with microscope,in the cells treated with drugs the adherence ability was reduced with refraction,shrink,circular,karyopycnosis,nuclear fragmentation and the cell chromatin agglutination.The apoptotic cells of solo drug groups were more than those in combined drug groups.MTT assay indicated that the MLT groups(0.5,1.0,2.5mmol·L-1)and MLT combined with DDP (25,50mg·L-1)groups all had inhibitory effects on the cell growth,and the inhibitory rate was increased with the concentrations of drugs.Two drugs had synergetic effect,CDI<1.Immunocytochemical staining results showed that the expressions of VEGF in solo drug groups and combined groups were lower than that in control groups,the expression of VEGF in combined groups was decreased significantly(P<0.05).Conclusion MLT can restrain the growth of ovarian cancer cells. MLT combined with DDP has synergetic effect on down-regulation of the expression of VEGF.Combined DDP with debita spissitudine MLT can achieve good curative effect.