丁酸钠诱导人慢性髓系白血病K562细胞肺耐药相关基因蛋白的表达  

Sodium Butyrate-Induced Expression of Lung Resistance-Related Gene in Human Myeloid Leukemia K562 Cells

作　　者：李宁[1] 钱新华[1] 姚英民[1] 王志远[2] 

机构地区：[1]第一军医大学附属南方医院儿科,广东广州510515 [2]第一军医大学生物医学工程系,广东广州510515

出　　处：《癌症》2003年第8期821-825,共5页Chinese Journal of Cancer

基　　金：广州市科技计划项目(No.2002J1-C0031)

摘　　要：背景与目的:肺耐药相关蛋白(lungresistance-relatedprotein,LRP)的过度表达是急性白血病患者对化疗不敏感及提示不良预后的指征之一。本研究探讨丁酸钠(sodiumbutyrate,NaB)对人慢性髓系白血病K562细胞LRP表达的诱导作用,并初步探讨LRP对细胞内阿霉素(adriamycin,ADM)、柔红霉素(daunorubicin,DNR)浓度的影响。方法:以K562细胞为体外模型,采用半定量逆转录聚合酶链反应(RT-PCR)检测NaB作用前后K562细胞LRPmRNA的水平;采用流式细胞仪结合间接免疫荧光检测比较NaB处理前后K562细胞LRP蛋白表达的变化。分别以流式细胞仪和荧光显微镜检测NaB处理前后K562细胞内DNR的蓄积和细胞内ADM分布的变化。结果:经2mmol/LNaB处理后较之处理前:①K562细胞的LRPmRMA水平明显增加;②LRP蛋白表达由阴性转为阳性,阳性细胞百分率由1.65%上升到35.81%;③经NaB处理后K562细胞内DNR的蓄积明显减少,细胞内DNR平均荧光较处理前减少了68.36%;④ADM在细胞核内分布明显减少,在细胞浆内分布明显增多。结论:NaB可诱导K562细胞LRPmRNA水平和蛋白表达的增加,LRP表达可造成细胞内DNR蓄积减少和ADR由细胞核向细胞浆转移。BACKGROUND &OBJECTIVE:Overexpression of lung resi stance related protein(LRP) is involved in multidrug resistance and poor outcome in acute leukemia. This study was designed to investigate the effect of sodium butyrate (NaB) on LRP expression level and the function of LRP in K562 cells. METHODS: Human myeloid leukemia K562 cells, used as an in vitro model, were treated with NaB. The LRP mRNA expression and protein levels in the cells before and after NaB treatment were detected by semi quantitative reverse transcription polymerase chain reaction (RT PCR) and flow cytometry indirect immunofluorescence method,respectively.Intracellular Adriamycin(ADM) location and intracellular daunorubicin(DNR) accumulation were detected by fluorescence microscope and flow cytometry, respectively. RESULTS: Compared with untreated K562 cells, the LRP mRNA level in the cells treated with 2 mmol/L NaB was increased obviously; the protein expression were turned from negative result to positive result and the ratios of positive cells were increased from 1.68%to 35.81%. Intracellular DNR accumulation was decreased in the treated K562 cells and the mean fluorescence of DNR reduced 68.64%compared with the untreated K562 cells. There was strong ADM fluorescence in the nuclei of untreated K562 cells; while the treated K562 cells showed significantly less ADM fluorescence in their nuclei but more fluorescence in the cytoplasm. CONCLUSION: Both the mRNA level and the protein expression of LRP in K562 cells can be increased by NaB induction. LRP induced by NaB is involved in the decreasing anti cancer drug accumulation and transporting the drugs from the nucleus to the cytoplasm in K562 cells.

关 键 词：丁酸钠 诱导 慢性髓系白血病 K562细胞 肺耐药相关基因蛋白 表达 

分 类 号：R733.72[医药卫生—肿瘤]