【摘要】 目的探讨氯化两面针碱对人口腔鳞癌KB细胞周期和凋亡的影响及相互关系。方法应用MTT比色法测定氯化两面针碱对细胞增殖的抑制作用;流式细胞术(FCM)检测氯化两面针碱对细胞周期的影响;Hoechst33258检测细胞凋亡;透射电镜观察细胞超微结构。结果氯化两面针碱在体外呈浓度依赖性显著抑制KB细胞的生长,48 h IC50为(2.36±0.22)μg/ml;与空白组相比,经3μg/ml氯化两面针碱作用, G2/M期细胞比例显著增多;6 μg/ml时凋亡细胞比例最高,可见典型的核染色质凝集、碎裂;9 μg/ml时,细胞凋亡率明显下降,同时镜下可见大量坏死细胞。结论 氯化两面针碱抗KB细胞的方式与其剂量密切相关,低浓度时以G2/M期阻滞为主,中浓度时诱导凋亡,高浓度时则致其坏死。
【关键词】 氯化两面针碱 G2/M期阻滞 凋亡 坏死 KB细胞
Abstract:ObjectiveTo explore the influence of nitidine chloride on cell cycle and apoptosis of human carcinoma of mouth floor KB cells and their relationship.MethodsThe proliferation inhibition rate and IC50(48 h) of nitidine chloride to KB cells were examined by MTT assay. The influence of nitidine chloride on cell cycle was measured by flow cytometry. Hoechst33258 technology was adopted to detect cell apoptosis. Transmission electron microscope was used to observe cell ultrastructure.ResultsNitidine chloride markedly suppressed proliferation of KB cells in a dose-dependent manner in vitro, IC50 for 48h was (2.36±0.22)μg/ml. Compared with control group, after treated with 3μg/ml of nitidine chloride,the percentage of G2/M phase cells significantly increased.The highest apoptotic percentage and typical nuclear chromatine condensation and fragmentation were observed at the nitidine chloride concentration of 6 μg/ml.When KB cells were treated with nitidine chloride of 9 μg/ml,cell apoptosis rate significantly decreased , many necrotic cells were seen under transmission electron microscope at the same time. ConclusionThe means of nitidine chloride inhibiting KB cells is closely associated with its dose,nitidine chloride induced G2/M phase arrest at low concentration, induced apoptosis at medium concentration and lead to necrosis at high concentration.
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