上海交通大学学报（医学版）

›› 2009, Vol. 29 ›› Issue (9): 1035-.

• 论著（基础研究） • 上一篇    下一篇

和厚朴酚对U937/ADR细胞系耐药逆转作用及其机制研究

薛 芳1, 成志勇2, 梁文同2, 陈 浩1, 王素云1, 姚 丽1, 潘 崚1   

  1. 1. 河北医科大学 第二医院血液内科 河北省血液病重点实验室, 石家庄 050000；2. 保定市第一医院血液肿瘤科, 保定 071000
  • 出版日期:2009-09-25 发布日期:2009-09-29
  • 通讯作者: 潘崚, 电子信箱：lingpan20002000@yahoo.com.cn。
  • 作者简介:薛芳（1972—）, 女, 主治医师, 博士；电子信箱：lovecell2006@gmail.com。
  • 基金资助:

    河北省科技攻关计划项目（072761130）

Resistance reverse effects of honokiol on multidrug resistance of U937/ADR cell line

XUE Fang1, CHENG Zhi-yong2, LIANG Wen-tong2, CHEN Hao1, WANG Su-yun1, YAO Li1, PAN Ling1   

  1. 1. Department of Hematology, Key Laboratory of Hematology of Hebei Province, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, China；2. Department of Hematology and Oncology, the First Hospital of Baoding, Baoding 071000, China
  • Online:2009-09-25 Published:2009-09-29
  • Supported by:

    Program of Scientific Problem Tackling of Hebei Province, 072761130

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摘要：

目的 探讨和厚朴酚（HNK）对白血病细胞U937阿霉素（ADR）耐药细胞系U937/ADR多药耐药逆转作用及其机制。方法 以大剂量（IC50）ADR短时间诱导方法，构建U937/ADR细胞系。以低浓度HNK（IC20）与不同化疗药物联合作用于U937/ADR细胞系，检测其对不同化疗药物耐药逆转倍数。罗丹明123检测药物外排功能；荧光定量PCR（FQ-PCR）和Western blotting检测不同浓度HNK对U937/ADR细胞系核转录因子-κB(NF-κB)和耐药相关基因MDR1及其蛋白P-糖蛋白（P-gp）表达的影响；ELISA法检测NF-κB亚单位p65（NF-κB p65）的DNA结合活性。结果 成功构建了白血病多药耐药细胞系U937/ADR，对ADR耐药指数为亲代U937细胞的11倍。6.5 μg/mL HNK可以逆转U937/ADR对ADR的耐药，逆转倍数为2.20倍。HNK能够浓度依赖性地下调U937/ADR细胞NF-κB、MDR1 mRNA和P-gp表达，抑制NF-κB p65的活性。结论 HNK能有效逆转U937/ADR多药耐药，其机制可能与抑制NF-κB p65活性、下调MDR1和P-gp表达有关。

关键词: 和厚朴酚, 多药耐药, U937/ADR, 核转录因子-κB, P-糖蛋白

Abstract:

Objective To investigate reverse effects of honokiol (HNK) on human multidrug resistance of in vitro leukemia U937/ADR cell line. Methods According to high-dose (IC50) short-term adriamycin induction method, human acute myeloid leukemia (AML) multidrug resistant U937/ADR cell line was produced. The drug resistances to different chemotherapeutic drugs were measured. Low-dose (IC20) HNK and different chemotherapeutic drugs were used to U937/ADR for measuring the drug resistance reversal fold. Drug excretion was tested by rhodamine 123 (Rho123). NF-κB (p65) and MDR1(P-gp) were detected by real-time fluorescent relativequantification reverse transcriptional PCR (FQ-PCR) or Western blotting after U937/ADR treated with different concentrations of HNK. NF-κB activity was also detected by NF-κB activity detection kits. Results Multidrug resistance U937/ADR cell line was successfully established and the ADR resistant index was 11 fold than U937 cells. Low concentration of HNK combination with adriamycin could reverse drug resistance and reversal index of ADR was 2.2 fold after treated with 6.5 μg/mL HNK. It also could decrease the NF-κB (p65) and MDR1(P-gp) expressions dose-dependently. Conclusion HNK could inhibit NF-κB activity to inhibit P-gp expression and reverse the multidrug resistance of U937/ADR cells.

Key words: honokiol, multidrug resistance, U937/ADR cell line, NF-κB, P-gp

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