高级搜索

多西他赛诱导肺腺癌A549/DTX细胞株的建立及耐药机制分析

刘晓冬, 常 青, 赵秀芳, 于 航, 侯毅鞠

刘晓冬, 常 青, 赵秀芳, 于 航, 侯毅鞠. 多西他赛诱导肺腺癌A549/DTX细胞株的建立及耐药机制分析[J]. 肿瘤防治研究, 2014, 41(06): 519-522. DOI: 10.3971/j.issn.1000-8578.2014.06.002
引用本文: 刘晓冬, 常 青, 赵秀芳, 于 航, 侯毅鞠. 多西他赛诱导肺腺癌A549/DTX细胞株的建立及耐药机制分析[J]. 肿瘤防治研究, 2014, 41(06): 519-522. DOI: 10.3971/j.issn.1000-8578.2014.06.002
LIU Xiaodong, CHANG Qing, ZHAO Xiufang, YU Hang, HOU Yiju. Establishment of Docetaxel Resistant Variant of Human Lung Adenocarcinoma Cell Line A549/DTX and Its Resistance Mechanism[J]. Cancer Research on Prevention and Treatment, 2014, 41(06): 519-522. DOI: 10.3971/j.issn.1000-8578.2014.06.002
Citation: LIU Xiaodong, CHANG Qing, ZHAO Xiufang, YU Hang, HOU Yiju. Establishment of Docetaxel Resistant Variant of Human Lung Adenocarcinoma Cell Line A549/DTX and Its Resistance Mechanism[J]. Cancer Research on Prevention and Treatment, 2014, 41(06): 519-522. DOI: 10.3971/j.issn.1000-8578.2014.06.002

多西他赛诱导肺腺癌A549/DTX细胞株的建立及耐药机制分析

基金项目: 吉林省教育厅科学技术研究项目(2012338)
详细信息
    作者简介:

    刘晓冬(1972-),男,本科,副主任医师,主要从事呼吸系统肿瘤研究

    通讯作者:

    侯毅鞠,E-mail:houyiju@163.com

  • 中图分类号: R734.2

Establishment of Docetaxel Resistant Variant of Human Lung Adenocarcinoma Cell Line A549/DTX and Its Resistance Mechanism

  • 摘要: 目的 建立人肺腺癌细胞A549多西他赛(DTX)耐药细胞株,并对其耐药机制进行初步分析。方法 采用逐步增加多西他赛浓度、间歇诱导的方法,建立人肺癌A549/DTX体外耐药细胞模型;MTT法检测A549/DTX的耐药特性;流式细胞仪比较多西他赛对耐药细胞株A549/DTX及亲本细胞株A549凋亡的影响;Western blot分析多西他赛作用后两种细胞凋亡调节因子Bc1-2、Bax表达差异。结果 MTT显示A549/DTX的多西他赛耐药指数为18.5;流式细胞分析显示经12.5、25、50 μg/L 多西他赛作用24 h后,A549/DTX的细胞凋亡率为(6.2±4.1)%、(13.6±2.7)%、(20.5±5.1)%,A549细胞凋亡率为(15.7±3.5)%、(28.5±2.9)%和(33.1±4.8)%,两者比较差异有统计学意义(P<0.01);Western blot显示,与A549相比,A549/DTX细胞Bcl-2蛋白表达明显升高,Bax蛋白明显降低(P <0.01)。结论 逐步增加浓度、间歇诱导的方法建立了稳定、耐药性较高的A549/DTX细胞株,并认为凋亡受抑是A549/DTX的耐药机制之一,与Bcl-2表达上调、Bax表达下调相关。

     

    Abstract: Objective To establish a human lung adenocarcinoma cell line A549 with the characterization of docetaxel (DTX) resistance, and investigate its biological mechanism of drug resistance. Methods A DTX resistant human lung adenocarcinoma cell line A549/DTX was obtained discontinuously by gradually increasing doses of DTX. The drug resistance of A549/DTX was evaluated by MTT assay. Apoptosis rates of DTX resistant human lung adenocarcinoma cell line A549/DTX and parental cell line A549 were detected by fl ow cytometry (FCM). The expression levels of Bc1-2 and Bax protein of the two cel1 1ines induced by different concentrations of DTX were detected by Western blot. Results Resistance index(IR) of A549/DTX was 18.5. The apoptosis rates induced by 12.5, 25 and 50 μg/L of DTX were (6.2±4.1)%, (13.6±2.7)% and (20.5±5.1)% in A549/DTX cells, signifi cantly lower than that in A549 cells which were (15.7 ±3.5)%, (28.5±2.9)% and (33.1±4.8)% respectively(P<0.01). Western blot showed a statistically higher expression of Bc1-2 and lower expression of Bax in A549/DTX than that in A549 cells at protein levels (P<0.01) . Conclusion A drug-resistant cell line A549/DTX with high IR was established by discontinuously induction and gradually increasing doses of DTX. Apoptosis inhibition is one of the mechanisms underlying the drug resistance of human lung adenocarcinoma cell line A549/DTX and is related with the up-regulation of Bc1-2 expression and down-regulation of Bax expression.

     

  • [1] Lv PT,Xiao J,Zhou K,et al.Mechanism research on the reversion of multidrug resistance in lung carcinoma cells A549 treated by coix lacryma-Jobi injection[J]. Zhongguo Quan Ke Yi Xue,2010,13(26):2956-8.[吕品田,萧娟,周坤,等.薏苡仁注射液 对肺腺癌细胞多药耐药性逆转作用的机制研究[J].中国全科医 学,2010,13(26):2956-8.]
    [2] Chen LB.Research on the mechanisms of docetaxel-resistant human lung adenocarcinoma[J].Zhongguo Zhong Liu Sheng Wu Zhi Liao Za Zhi,2012,19(5):467-71.[陈龙邦.人肺腺 癌多西他赛耐药机制的研究[ J ] . 中国肿瘤生物治疗杂 志,2012,19(5):467-71.]
    [3] Tsavaris N,Kosmas C,Skopelitis E,et al.A phaseⅡstudy of the docetaxel-carboplatin chemotherapy regimen in advanced nonsmall- cell lung cancer[J].Lung,2005,183(6):405-16.
    [4] Sun H,Geng J, Chen LB. Research development on drug-resistance mechanism of taxanes[J]. J Med Postgrad,2007,20(3):315-8,323. [孙海,耿建,陈龙邦.紫杉类药物耐药机制的研究进展[J].医学研 究生学报,2007,20(3):315-8,323.]
    [5] Zhang N,Xiong YY,Li L. The relationship between expression of survivin, bcl-2, bax and multidrug resistance gene in non-small cell lung cancer[J].Zhong Liu Fang Zhi Yan Jiu,2006,33(3):168-70.[ 张娜,熊永炎,李莉.非小细胞肺癌中 survivin、bcl-2和bax蛋白与多药耐药基因的表达及其相关性 [J] 肿瘤防治研究,2006,33(3):168-70.]
    [6] Del Bufalo D, Biroccio A, Trisciuoglio D, et al.Bcl-2 has differing effects on the sensitivity of breast cancer cells depending on the antineoplastic drug used[J].Eur J Cancer,2002,38(18):2455-62.
    [7] Ji FY, Qian GS, Qian P, et al. Involvement of bcl-2 in multidrug resistance in human small cell lung cancer cell subline H446/ DDP[J]. Zhonghua Jie He He Hu Xi Za Zhi,2006,29(3):156-60. [ 戢福云, 钱桂生, 钱频, 等. B c l - 2 对人小细胞肺癌细胞系 H446/DDP多药耐药性的影响[ J ] . 中华结核和呼吸杂 志,2006,29(3):156-60.]
    [8] Hu Y,Bebb G,Tan S,et al.Antitumor efficacy of oblimersen Bcl-2 antisense oligonucleotide alone and in combination with vinorelbine in xenograft models of human non-small cell lung cancer[J]. Clin Cancer Res,2004,10(22):7662-70.
    [9] Ghobrial IM,Witzig TE,Adjei AA.Targeting apoptosis pathways in cancer therapy[J]. CA Cancer J Clin,2005,55(3):178-94.
    [10] Jendrossek V.The intrinsic apoptosis pathways as a target in anticancer therapy[J].Curr Pharm Biotechnol,2012,13(8)1426-38.
    [11] Sotiropoulou PA,Candi A,MascréG,et al.Bcl-2 and accelerated DNA repair mediates resistance of hair follicle bulge stem cells to DNA-damage-induced cell death[J].Nat Cell Biol,2010,12(6):572-82.
计量
  • 文章访问数:  1914
  • HTML全文浏览量:  324
  • PDF下载量:  1074
  • 被引次数: 0
出版历程
  • 收稿日期:  2013-04-15
  • 修回日期:  2013-09-20
  • 刊出日期:  2014-06-24

目录

    /

    返回文章
    返回
    x 关闭 永久关闭