【作者】 薛建秀；

【导师】 王斌全；

【作者基本信息】 山西医科大学 ， 耳鼻咽喉科学， 2004， 硕士

【摘要】 目的：观察三氧化二砷(As2O3)对喉癌Hep-2细胞株的作用和对细胞周期的影响，为抗肿瘤药物的筛选和用药式提供理论依据。方法：体外培养的Hep-2细胞与不同浓度的As2O3连续作用24、48、72h,用倒置显微镜、流式细胞仪研究细胞凋亡的形态学改变，检测细胞凋亡率，进行细胞周期分析。DNA琼脂糖凝胶电泳定性研究细胞凋亡。结果：倒置显微镜观察发现, As2O3诱导Hep-2细胞悬浮细胞增多，贴壁细胞形状变圆、体积增大，细胞增殖减慢具有时间和药物浓度双相依赖性。流式细胞仪检测发现不同药物浓度和作用时间下均有细胞凋亡发生，相关性分析显示药物浓度和时间与凋亡发生呈正相关,即药物浓度增加细胞凋亡增加,随着作用时间延长凋亡发生也增加。2μmol/L As2O3作用48h、72hDNA琼脂糖凝电泳呈现明显的梯度带, 流式细胞仪图中出现典型的“亚G1期”峰，定性定量证明As2O3可诱导Hep-2细胞凋亡。As2O3能造成细胞周期明显变化，2μmol/L As2O3作用24h时,S期细胞比例增高,72h后,S期细胞明显下降,细胞大量凋亡。As2O3在作用早期,阻滞细胞通过S期,随着时间的延长,诱导S期细胞凋亡。结论：As2O3可有效抑制人喉癌Hep-2细胞的体外生长,具有时间浓度依赖性特点。As2O3诱导Hep-2细胞凋亡主要是S期的细胞。更多还原

【Abstract】 Objective: To observe the apoptosis and the cell cycle in human laryngeal squamous cell carcinoma strain Hep-2 exposed to arsenic trioxide (As2O3). Methods:Using flow cytometry ,light microscopy and DNA agarose gel electrophoresis technique, we investigated in vitro Hep-2 cells in different conditions.Results: Hep-2 was adherent cells in normal survival condition. As2O3 affected Hep2 cell growth apparently. Under fluorescence microscope,necrotic cells were red, normal cells were evenly blue ,and apoptotic cells were blue with condensed and fragmented nuclei.The latter and the increased volume of cells appeared to be time and dose dependent. DNA”ladder” and obviousSub-G1 phase peak occurred in flow cytometry were observed for Hep-2 cell exposed to 2μmol/L As2O3 48h and 72h .The most prominent effect As2O3 of on cell cycle kinetics was a slowdown in S-phase transit during which cells undergone apoptosis.S-phase cell percentage increased after exposed to for 24h .With time elapsing,S-phase cell reduced.Conclusion: As2O3 can induce apoptosis of Hep-2 cell,which can be related to the cell cycle arrest.更多还原