Li Ning, Qian Xin-Hua, Yao Ying-Min, Wang Zhi-Yuan
Department of Pediatrics, Nanfang Hospital, The First Military Medical University, Guangzhou, Guangdong, 510515, PR China.
Ai Zheng. 2003 Aug;22(8):821-5.
BACKGROUND & OBJECTIVE: Overexpression of lung resistance-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.
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.
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.
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.
肺耐药相关蛋白(LRP)的过表达与急性白血病的多药耐药及不良预后有关。本研究旨在探讨丁酸钠(NaB)对K562细胞中LRP表达水平及功能的影响。
以人髓系白血病K562细胞作为体外模型,用NaB处理。分别采用半定量逆转录聚合酶链反应(RT-PCR)和流式细胞术间接免疫荧光法检测NaB处理前后细胞中LRP mRNA表达及蛋白水平。分别用荧光显微镜和流式细胞术检测细胞内阿霉素(ADM)的定位及柔红霉素(DNR)的蓄积。
与未处理的K562细胞相比,2 mmol/L NaB处理的细胞中LRP mRNA水平明显升高;蛋白表达由阴性转为阳性,阳性细胞比例从1.68%增至35.81%。处理后的K562细胞内DNR蓄积减少,与未处理的K562细胞相比,DNR平均荧光降低68.64%。未处理的K562细胞核中有较强的ADM荧光;而处理后的K562细胞核中ADM荧光明显减少,细胞质中荧光增多。
NaB诱导可使K562细胞中LRP的mRNA水平及蛋白表达均增加。NaB诱导的LRP参与降低K562细胞中抗癌药物的蓄积,并将药物从细胞核转运至细胞质。