汉防己甲素干预K562细胞mdr1基因表达的研究  

Effect of tetrandrine on the doxorubicin-induced expression of mdrl gene in K562 cells

作　　者：吕旭晶[1] 许文林[1] 罗文娟[1] 王法春[1] 陈巧云[1] 

机构地区：[1]江苏大学附属人民医院血液科,镇江市212002

出　　处：《中华血液学杂志》2008年第7期468-471,共4页Chinese Journal of Hematology

基　　金：江苏省卫生重大课题基金（K2005017）

摘　　要：目的观察汉防己甲素（TTD）对阿霉素诱导的K562细胞mdr1基因表达的影响，并初步探讨其机制。方法采用MTT法观察TTD对K562细胞的毒性作用，以0．6 μg/ml阿霉素单独作用或0．6 μg/ml阿霉素联合不同浓度的TTD（0．5、1．0、2．0 μg/ml）作用于K562细胞，用RT—PCR法检测mdr1 mRNA、NF—κB mRNA水平，用流式细胞术检测P—gp的表达情况，用胞内罗丹明123（Rh0123）积聚试验检测P—gP的功能。结果空白对照K562细胞未见明显mdr1 mRNA及P-gP表达（水平分别为0．171±0．012、7．85±0．15），细胞内Rh0123平均荧光强度（反映P—gP功能）为711．9±63．6，NF—κB mRNA水平为0．783±0．090；0．6 μg／ml阿霉素作用24h后mdr1 mRNA、P—gP、NF—κB mRNA表达上调为0．428±0．012、73．68±1．84、1．075±0．047，细胞内Rh0123平均荧光强度下降为347．8±60．6（P值均〈0．05）；2．0 μg／ml TTD预作用24h再与0．6 μg／ml阿霉索联合作用24h能显著抑制阿霉素诱导的mdr1 mRNA、P—gP表达及功能、NF—κB mRNA的上调（分别为0．148±0．006、7．18±0．38、799．7±45．8、0．627±0．098）（P〈0．05），而0．5、1．0 μg／ml TTD无明显影响。结论TTD以浓度依赖性的方式干预阿霉素诱导的mdr1 mRNA、P—gP表达和P-gP功能的上调，其机制可能与TTD抑制了阿霉素诱导的NF—κB的表达有关。Objective To investigate the effect of tetrandrine （ TTD ） on doxorubicln-induced mdrl gene expression and its mechanism. Methods MTF assay was used to detect the cytotoxicity of TTD to K562 cells. K562 cells were treated with doxorubicin alone or 0.6 μg/ml doxorubicin combined with various concentrations of TTD. RT-PCR was used to detect the mRNA expression of mdrl and NF-κB. Flow cytometry was used to assay the expression of P-glycoprotein （P-gp）. Intracellular rhodamine 123 （Rho123） retention assay was applied to test the P-gp function. Results After treatment with 0.6 μg/ml doxorubicin for 24 hours, the expressions of mdrl mRNA, NF-KB mRNA and P-gp in K562 cells were increased from 0. 171 ± 0.012, 0. 783 ± 0. 090, 7.85 ± 0.15 to 0. 428 ± 0. 012,1. 075 ± 0. 047 and 73.68 ± 1.84, respectively. The intracellular Rho123 retention was decreased from 711.9 ± 63.6 to 347.8 ± 60.6, indicating up-regulation of P-gp function （ P 〈 0.05 ）. Pretreatment of K562 cells with 2.0 μg/ml TTD for 24 hours and then incubated for another 24 h with doxorubicin, the expressions of mdrl mRNA, NF-κB mRNA, P-gp and up-regulation of P-gp function induced by doxorubicin were prevented in K562 cells（0. 148 ± 0. 006, 0. 627 ± 0. 098,7.18 ± 0.38 and 799.7 ± 45.8, respectively P 〈 0.05 ）. But 0.5 μg/ml and 1.0 μg/ml TTD had little effect. Conclusions TTD inhibits the expression of mdr1 mRNA, P-gp and up-regulated P-gp function induced by doxorubicin in a dose dependent manner. The mechanism of this effect may be down-regulation of NF-κB by TTD.

关 键 词：K562细胞 汉防己甲素 MDR1基因 NF—κB 

分 类 号：R686[医药卫生—骨科学]