IL-12联合DDP对人卵巢癌HO-8910细胞周期及增殖的影响

Effect of IL-12 combined with DDP on cell cycle and proliferation of human oophoroma HO-8910 cells in vitro

目的:探讨IL-12联合化疗药物顺铂(DDP)对人卵巢癌HO-8910细胞周期及增殖的影响,并阐明其抑制肿瘤细胞增殖的机理。方法:取对数生长期的人卵巢癌HO-8910细胞株分为对照组、单独使用DDP组、单独使用IL-12组及IL-12联合DDP组。按作用时间(0、24、48、72、96、120 h)和DDP浓度(0、2、4、8、16、32μmol/L)不同,采用MTT比色法分别测定IL-12单独或联合应用DDP对人卵巢癌HO-8910细胞的影响,绘制细胞生长曲线和计算细胞增殖抑制率。根据细胞生长曲线评价肿瘤细胞的增殖速度,流式细胞术检测细胞周期及细胞凋亡的变化,倒置显微镜和电镜观察细胞形态学改变。结果:单独应用DDP或IL-12时,其抑制率在96 h分别为28.93%和29.12%,而IL-12联合DDP在48 h抑制率为27.45%。HO-8910、HO-8910+DDP和HO-8910+IL-12细胞生长抑制曲线随时间延长而升高,在第96 h开始升高(P<0.01),而加入IL-12+DDP细胞的生长抑制曲线则从第48 h就开始升高,72 h达高峰(P<0.05);流式细胞仪检测显示,处于G1/G2期的HO-8910+IL-12+DDP细胞明显减少,而G2/M、S期细胞明显增多;电镜结果显示,HO-8910+IL-12+DDP细胞存在内质网扩张,线粒体空泡化,溶酶体增生,部分细胞可见染色质边集等凋亡前期改变。结论:IL-12联合DDP能抑制人卵巢癌HO-8910细胞增殖,有效地诱导细胞凋亡,增强DDP对肿瘤的治疗作用,为临床卵巢癌患者的治疗提供理论依据。

Objective：To explore the effect of interleukin-12（IL-12） combined with DDP on the cell cycle and proliferation of human oophoroma HO-8910 cells,expound the mechanisms.Methods：HO-8910 cells at logarithmic growth phase were divided into control group,DDP group,IL-12 group and IL-12＋DDP group.According to different response times（0,24,48,72,96,120 hours） and different concentrations of DDP（0,2,4,8,16,32 μmol/L）,the effects of IL-12 or IL-12＋DDP on human oophoroma HO-8910 cells were detected by MTT assay.The cell growth curve was drew and the inhibitory rate of cell proliferation was calculated.The cell proliferation speed was evaluated according to cell growth curve,the changes of cell cycle and cell apoptosis were detected by flow cytometry,and the morphological changes of HO-8910 cells were observed by invert microscope and transmission electron microscope.Results：The inhibition rates of DDP and IL-12 were 28.93% and 29.12% at 96 hours,respectively;the inhibition rate of IL-12＋DDP was 27.45% at 48 hours.The cell growth inhibition curve in HO-8910,HO-8910＋DDP and HO-8910＋IL-12 groups raised at 96 hours with time（P〈0.01）.However,the cell growth inhibition curve in HO-8910＋IL-12＋DDP group begin to raise at 48 hours and reached peak at 72 hours（P〈0.05）;flows cytometry analysis showed that in G1/G2 phase,the cells of HO-8910＋IL-12＋DDP group decreased obviously,while in S phase and G2/M phase,cells increased remarkably;electron microscope results showed that HO-8910＋IL-12＋DDP cells existed apoptotic prophase changes,such as endoplasmic reticulum dilatation,plastochondria vacuole,lysosome proliferation,obvious chromatin collection in partial cells.Conclusion：IL-12 combined with DDP can suppress the proliferation of human oophoroma HO-8910 cells,induce cell apoptosis effectively,enhance the therapeutic effect of DDP on oophoroma and provide a scientific basis for the treatment of oophoroma.