rmhTRAIL诱导耐阿霉素细胞株K562/A02凋亡及作用机制的实验研究  

The mechanism of the apoptosis of the leukemia cell line K562 resistant to Adriamycin K562/A02 induced by rmhTRAIL

作　　者：王雅茹[1] 王福旭[2] 温树鹏[2] 杜行严[2] 杨渤彦[3] 张学军[2] 杨世方[4] 

机构地区：[1]保定市第三医院肿瘤血液科,河北保定071000 [2]河北医科大学附属二院血液科,河北石家庄050000 [3]中国医学科学院协和医科大学肿瘤医院,北京100029 [4]北京沙东生物技术有限公司,北京100081

出　　处：《现代肿瘤医学》2008年第5期713-718,共6页Journal of Modern Oncology

摘　　要：目的:探讨recombinanat mutant human TNF-related apoptosis -inducing ligand,(rmhTRAIL)对阿霉素诱导的人慢性粒细胞白血病红白血病变耐药细胞株K562/A02(mdr-1+)的促凋亡作用及其机理。方法:以亲本阿霉素敏感细胞株K562(mdr-1-)为对照,观察rmhTRAIL作用后K562、K562/A02细胞形态变化,采用碘化丙啶染色法流式细胞术(FACSCalibur)定量检测细胞sub -G1%, MTT法测定rmhTRAIL对细胞增殖抑制,半定量逆转录-聚合酶链反应(semiquantitative reverse transcription polymerase chain reaction,RT-PCR)方法检测K562、K562/A02细胞的四种TRAIL受体DcR1、DcR2、DR4、DR5mRNA表达水平。结果:rmhTRAIL可诱导K562、K562/A02细胞凋亡,有典型的细胞形态改变;流式细胞术检测显示K562/A02的sub -G1%大于K562 (P<0.05); rmhTRAIL对K562/A02的增殖抑制作用优于K562 (P<0.05); K562/A02中DR4、DR5mRNA的表达高于K562, DcR1mRNA在K562/A02表达低于K562, DcR2 mRNA在两种细胞中均不表达。结论:rmhTRAIL可以诱导K562/A02细胞凋亡;rmhTRAIL对K562/A02促凋亡和抑制增殖作用优于其亲本细胞K562;K562/A02细胞表面TRAIL死亡受体高表达及诱骗受体低表达可能是耐药细胞更敏感的原因。Objective：To investigate the apoptosis mechanism of multi - drug resistant cell line K562/A02 （ mdr- 1^＊） induced by rmhTRAIL. Methods： K562 （ mdr - 1^- ） was controlled, morphology was observed before and after treatment of rmhTRALL; Flow cytometer was used to analyze the apoptotic peak of various concentrations of rmhTRAIL; MTT to assay the growth inhibitory effect of rmhTRAIL; The different mRNA expression levels of DR4, DRS, DcR1, DcR2 of K562 and K562/A02 cells were examined by semi - quantitative reverse transcription polymerase chain reaction RT - PCR. Results： The apoptosis of K562, K562/A02 cells induced by rmhTRAIL was observed at a cytomorphology angle,with typical cytmorphological changes; The apoptosis ratio of K562/A02 group surpassed K562 （P 〈 0.05 ） ; The inhibition of K562/A02 cells surpassed the K562 （ P 〈 0.05） ; K562/A02 ceils expressed DR4 mRNA and DR5 mRNA higher than K562, the expression of DcR1 on K562/A02 cells was lower than K562, DcR2 mRNA did not express in two kind of cells. Conclusion： rmhTRAIL can induce K562 and K562/A02 cells apoptosis; The efficacy of rmhTRAIL to induce apoptosis and inhibit proliferation in K562/A02 cell line is more predominant than their parental cell line in the study ;The high expression of death receptors and the low expression of decoy receptors could be one of the reasons that K562/A02 cells were more sensitive than K562.

关 键 词：肿瘤坏死因子相关凋亡诱导配体 多药耐药基因 K562 K562/A02 凋亡 机制 死亡受体 

分 类 号：R73-3[医药卫生—肿瘤] R733.7[医药卫生—临床医学]