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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 investigate the effect of DDP on the cell cycle and proliferation of human ovarian cancer cell line HO-8910 and to clarify its mechanism of inhibiting tumor cell proliferation. Methods: HO-8910 cell line of human ovarian cancer was divided into control group, DDP group alone, IL-12 group and IL-12 combined with DDP group alone. According to the different action time (0,24,48,72,96,120 h) and the concentration of DDP (0,2,4,8,16,32μmol / L), MTT assay was used to determine IL-12 alone or in combination Application of DDP on human ovarian cancer HO-8910 cells, plot cell growth curve and calculate the cell proliferation inhibition rate. The proliferation rate of tumor cells was evaluated according to cell growth curve. The cell cycle and apoptosis were detected by flow cytometry. The morphological changes of cells were observed by inverted microscope and electron microscope. Results: When DDP or IL-12 alone was used, the inhibition rates were 28.93% and 29.12% at 96 h, respectively. The inhibition rate of IL-12 and DDP at 48 h was 27.45%. The growth inhibition curves of HO-8910, HO-8910 + DDP and HO-8910 + IL-12 cells increased with time and began to increase at 96 h (P <0.01) The growth inhibition curve increased from the 48th hour to the peak at 72 hours (P <0.05). Flow cytometry showed that the number of HO-8910 + IL-12 + DDP cells in G1 / G2 phase decreased significantly G2 / M, S phase cells were significantly increased; electron microscopy showed that HO-8910 + IL-12 + DDP cells exist endoplasmic reticulum dilation, mitochondria vacuolization, lysosomal hyperplasia, some cells visible chromatin edge set of apoptosis Early changes. Conclusion: IL-12 combined with DDP can inhibit the proliferation of human ovarian cancer cell line HO-8910, induce cell apoptosis effectively and enhance the therapeutic effect of DDP on the tumor, so as to provide a theoretical basis for the treatment of clinical ovarian cancer patients.