【作者】 刘景；

【导师】 丁剑午；

【作者基本信息】 南昌大学 ， 肿瘤学， 2011， 硕士

【摘要】 目的：通过本实验观察全反式维甲酸(Alltrans Retinoic Acid,ATRA)、奈达铂(Nedaplatin,NDP)及其两药联合应用对人肝癌细胞株Huh-7增殖与凋亡的影响,探讨本实验中化疗药物联合应用的相互作用。方法：通过选取不同的实验浓度如下,ATRA分设10-6、10-5、10-4mol/L三组、NDP设1、2、5mg/L三组,随后通过预实验筛选出ATRA10-5mol/L联合不同浓度NDP(1、2、5mg/L)分别作用于肝癌Huh-7细胞24、48、72h；另外,将ATRA(10-5 mol/L)+NDP(1mg/L)作为联合处理药物组作用于肝癌Huh-7细胞24h、48h、72h；应用医用倒置显微镜观察实验前后HUh-7细胞的形态学变化,四甲基偶氮唑蓝(MTT)法观察ATRA、NDP及其两药联合应用对肝癌Huh-7细胞增殖的影响；同时采用流式细胞仪分析其对肝癌Huh-7细胞凋亡率的影响。结果：实验中不同浓度的全反式维甲酸ATRA(10-6、10-5、10-4 mol/L)和奈达铂NDP(1、2、5mg/L)单独作用于肝癌Huh-7细胞均有明显的抑制作用,且该作用随时间的延长、浓度的增高也渐增强；ATRA与NDP两药联合应用于HUh-7细胞后表现为协同作用,联合用药组与单药组相比,P<0.01。用流式细胞仪进行凋亡率检测,结果显示：ATRA处理HUh-7细胞48h后的凋亡率为：对照组0.82±0.15%, ATRA(10-6mol/L)组31.9±1.91%,ATRA(10-5mol/L)组42.57±1.03%,实验组与对照组比较(P<0.05)；NDP单药处理48h后,对照组为0.34±0.07%,药物组即1、2、5mg/L凋亡率分别为28.49±0.6%、38.53±0.7%、46.6±1.1%；ATRA、NDP两药联合作用48 h后,对照组为0.82%±0.15%,NDP 1 mg/L组为28.49%±0.6%,ATRA 10-5 mol/L组为42.57%±1.03%, ATRA10-5mol/L+NDP 1mg/L联合作用组为55.35%±1.30%,与单药比较差异有显著统计学意义(P<0.01)。结论：全反式维甲酸ATRA与奈达铂NDP对人肝癌Huh-7细胞株均有抑制增殖和诱导凋亡的作用,且联合用药组与单药组比较,全反式维甲酸ATRA与奈达铂NDP两药联合应用于肝癌Huh-7细胞,具有明显协同作用。更多还原

【Abstract】 Objective:By this experiment, to observed the effects of Alltrans Retinoic Acid (ATRA) and Nedaplatin (NDP) alone and their combined effects for cell proliferation and apoptosis on human hepatoma cell line Huh-7 in vitro, and to investigated the effect combination of chemotherapy medicine to cure hepatoma.Methods:By choosing the medicines at different concentrations of ATRA(10-6、10-5 10-4mol/L) and NDP (1、2、5mg/L)alone and combine effects for hepatoma cell 24、48、72h, Others, the medicines effective final concentration determined by pre-screen test, ATRA(10-5mol/1) combined with NDP (1mg/1) was used to treat hepatoma cells for 24、48、72h. Cell morphology changes was detected using inverted micros-copy; MTT-test was used to estimate the inhibition of proliferation on hepatoma cell line Huh-7;flow cytometry was applied to analyze the effect of cell apoptosis.Results:Experiment with different concentrations of ATRA(10-6,10-5,10-4 mol/1) and NDP (1、2、5mg/1)could markedly inhibit the cell proliferation,and the effect depended on exposure time and concentration, and the effect of combining was additive and synergistic, which had statistical significance comparing with ATRA and Nedaplatin alone(P<0.01).Apoptosis rate by flow cytometry, and the results showed, ATRA effected Huh-7 cells apoptosis rate(48h):control group 0.82±0.15%, ATRA(10-6mol/L) 31.9±1.91%, ATRA (10-5mol/L) 42.57±1.03%, comparing with the experiment group and control group (P<0.05); NDP effected Huh-7 cells apoptosis rate(48h):the control group 0.34±0.07%, the drug groups of NDP(1、2、5 mg/L) were:28.49±0.6%,38.53±0.7% and 46.6±1.1%; and the combined use resulted in synergic effect, NDP (1 mg/1), ATRA (10-5ol/1) and the two combined,they induce the apoptosis rate of Huh-7 at 48h were 28.49%±0.6%、42.57%±1.03%、55.35%±1.30%. So that the combined group comparing with the single groups, P<0.01.Conclusion:Both ATRA and NDP can inhibit cell proliferation and promote cell apoptosis of human hepatoma cell line Huh-7,and comparing with the single groups,ATRA combined with NDP can significantly increase the chemosensitivity on hepatoma cells.更多还原