冬凌草甲素对神经母细胞瘤抗肿瘤的作用研究
投稿时间:2015-12-18  修订日期:2015-12-19  点此下载全文
引用本文:张李迪,高丰厚,姜斌.冬凌草甲素对神经母细胞瘤抗肿瘤的作用研究[J].医学研究杂志,2016,45(8):28-32
DOI: 10.11969/j.issn.1673-548X.2016.08.009
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作者单位E-mail
张李迪 201999 上海交通大学医学院附属第三人民医院肿瘤科  
高丰厚 201999 上海交通大学医学院附属第三人民医院肿瘤科  
姜斌 201999 上海交通大学医学院附属第三人民医院肿瘤科 jiangbinwcr@sjtu.edu.cn 
基金项目:国家自然科学基金资助项目(81302006)
中文摘要:目的 在体内体外水平探讨冬凌草甲素(oridonin)对神经母细胞瘤的抗肿瘤作用。方法 CCK-8法检测冬凌草甲素对细胞增殖的影响;AO/EB双荧光染色后荧光显微镜观察细胞凋亡情况;免疫印迹检测细胞内活化的caspase-3和剪切的PARP表达水平;构建SHSY-5Y神经母细胞裸鼠异种移植瘤模型,Ki-67免疫组化染色分析冬凌草甲素对肿瘤组织增殖的影响。结果 冬凌草甲素作用HSY-5Y和SK-N-MC细胞24h后,可以剂量依赖性抑制细胞增殖。不同浓度冬凌草甲素处理24h后AO/EB双荧光染色显示,随着药物浓度增加细胞凋亡明显增多。免疫印迹结果证实冬凌草甲素可以浓度依赖性增加活化的caspase-3和剪切的PARP蛋白的表达。冬凌草甲素处理组裸鼠移植瘤体积增量显著小于对照组,Ki-67免疫组化显示冬凌草甲素处理组肿瘤组织增殖细胞比率较对照组显著降低。结论 冬凌草甲素可以抑制SHSY-5Y和SK-N-MC细胞增殖并诱导其凋亡,并在动物水平抑制裸鼠移植瘤的生长。
中文关键词:冬凌草甲素  神经母细胞瘤  SHSY-5Y细胞  SK-N-MC细胞  异种移植瘤
 
Antitumor Efficacy of Oridonin on Neuroblastoma
Abstract:Objective To investigate the antitumor efficacy of oridonin on neuroblastoma in vitro and in vivo. Methods The CCK-8 assay was used to measure the inhibitory effect of oridonin on proliferation of SHSY-5Y and SK-N-MC cells. Apoptosis was determined by fluorescence microscope after AO/EB staining. The expression of cleaved caspase-3 and cleaved PARP was determined by immunoblot analysis. BALB/c nude mouse xenograft models with neuroblastoma were built by SHSY-5Y cell line and the proliferation of tumor tissues were assessed by Ki-67 immunohistochemistry. Results Treatment with oridonin for 24h resulted in a marked decrease in cell viability of SHSY-5Y and SK-N-MC cells in a dose-dependent manner. After treatment with an increasing concentration of oridonin for 24h, the percentage of apoptotic cells observed by AO/EB staining increased. Treatment with oridonin increased the expression of cleaved caspase-3 and PARP. The tumor volume increment and the proliferation of tumor tissues measured by Ki-67 immunostaining in models with oridonin were significantly less than vehicle control. Conclusion Oridonin can exert its antitumor effect by inhibiting proliferation and inducing apoptosis of SHSY-5Y and SK-N-MC cells in vitro and in vivo.
keywords:Oridonin  Neuroblastoma  SHSY-5Y  SK-N-MC  Xenograft
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