洋地黄毒苷对肺癌NCI-H446与A549细胞增殖和周期的影响

doi:10.3969/j.issn.1004-616x.2015.01.002

癌变·畸变·突变

洋地黄毒苷对肺癌NCI-H446与A549细胞增殖和周期的影响

刘锋，陈志添，邵孔健，萨本仲

福州市第二医院肝胆外科，福建福州 350007

收稿日期:2014-07-14 修回日期:2014-10-21 出版日期:2015-01-31 发布日期:2015-01-31
作者简介:刘锋，E-mail：508788420@qq.com； Tel：18046041710
基金资助:
福州市科技计划项目(2011-S-67-4)

Effect of digitoxin on proliferation and cell cycle of NCI-H446 and A549 cells

LIU Feng，CHEN Zhitian，SHAO Kongjian，SA Benzhong

Department of Hepatobiliary Surgery, The Second Hospital of Fuzhou, Fuzhou 350007, Fujian, China

Received:2014-07-14 Revised:2014-10-21 Online:2015-01-31 Published:2015-01-31

摘要/Abstract

摘要：

目的：探讨洋地黄毒苷对肺癌NCI-H446与A549细胞增殖和细胞周期的影响，并初步探讨其可能的作用机制。方法：体外培养的NCI-H446与A549细胞分别经不同浓度(10、20、40、80、160 nmol/L)的洋地黄毒苷处理24、48和72 h后，采用MTT法检测细胞增殖情况，应用流式细胞术检测洋地黄毒苷处理细胞48 h后各组细胞周期分布，Western blot检测细胞中Cyclin A和P21蛋白表达水平。结果：与未经洋地黄毒苷处理的对照组相比，不同浓度(10、20、40、80和160 nmol/L)的洋地黄毒苷均可抑制 NCI-H446与A549细胞的增殖，均呈剂量和时间依赖性(P<0.05)，作用48 h后，洋地黄毒苷对NCI-H446与A549细胞的IC50值分别为61.26 nmol/L和110.73 nmol/L。流式细胞仪分析结果显示，随药物作用浓度增加，G0/G1细胞比例降低，S期细胞比例显著增加，与对照组相比，差异具有统计学意义(P<0.05)。Western blot分析结果显示，洋地黄毒苷能剂量依赖性的下调Cyclin A1蛋白表达和上调P21 蛋白的表达(P<0.05)。结论：洋地黄毒苷可抑制体外培养的肺癌NCI-H446与A549细胞增殖，诱导细胞发生S期阻滞，其机制可能与细胞周期相关调控蛋白的表达有关。

关键词: 洋地黄毒苷, 肺癌, 细胞周期调控, 细胞周期蛋白A1

Abstract:

OBJECTIVE: To investigate the effects of digitoxin on proliferation and cell cycle of human lung cancer cell lines NCI-H446 and A549，and to explore its possible mechanisms. METHODS：NCI-H446 and A549 cells were treated with digitoxin at different concentrations(10，20，40，80 and 160 nmol/L). The effect on proliferation of NCI-H446 and A549 cells were detected by MTT assay. Flow cytometry was used to test the cell cycle distribution of NCI-H446 and A549 cells. Protein expressions of Cyclin A1 and P21 in NCI-H446 and A549 cells were evaluated by Western blot. RESULTS：As compared with control group，cell proliferation was inhibited by digitoxin in a dose- and time-dependent manner(P＜0.05)，the IC50 value for digotoxin were 61.26 nmol/L in NCI-H446 and 110.73 nmol/L in A549 at 48 h. After 48 h treatment，the proportion of NCI-H446 and A549 cells in G0/G1 phase was decreased，while the proportion in S phase was increased. Western blot analysis showed that digitoxin could significantly up-regulate the expression of Cyclin A1 and down-regulate the expression of P21 in a dose-dependent way(P<0.05). CONCLUSION： Digitoxin could exert an the anti-proliferative effect on NCI-H446 and A549 cells and induce S phase arrest in vitro. The mechanism may be related to the expressions of proteins associated with cell cycle.

Key words: digitoxin, lung cancer, regulation of cell cycle, Cyclin A1

刘锋，陈志添，邵孔健，萨本仲. 洋地黄毒苷对肺癌NCI-H446与A549细胞增殖和周期的影响[J]. 癌变·畸变·突变, doi: 10.3969/j.issn.1004-616x.2015.01.002.

LIU Feng，CHEN Zhitian，SHAO Kongjian，SA Benzhong. Effect of digitoxin on proliferation and cell cycle of NCI-H446 and A549 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2015.01.002.

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洋地黄毒苷对肺癌NCI-H446与A549细胞增殖和周期的影响

Effect of digitoxin on proliferation and cell cycle of NCI-H446 and A549 cells

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