HIF-1α-PDK1信号系统参与急性单核细胞白血病糖代谢和耐药的机制研究  

Mechanism of HIF-1α-PDK1 signaling system involved in glucose metabolism and drug resistance in acute monocytic leukemia

作　　者：许晓巍[1] 赵初娴[1] 李肃[1] 王椿[1] 宋献民[1] 白海涛[1] XU Xiao-wei;ZHAO Chu-xian;LI Su;WANG Chun;SONG Xian-min;BAI Hai-tao(Department of Hematology,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200080,China)

机构地区：[1]上海交通大学附属第一人民医院血液科,上海200080

出　　处：《上海交通大学学报（医学版）》2018年第11期1294-1299,共6页Journal of Shanghai Jiao tong University:Medical Science

基　　金：上海市卫生和计划生育委员会科研课题面上项目(201440292)~~

摘　　要：目的·探讨HIF-1α-PDK1信号系统调控急性单核细胞白血病细胞糖代谢以及耐药的机制。方法·定量聚合酶链式反应(quantitative polymerase chain reaction,qPCR)检测U937细胞、U937/DNR细胞和原代急性单核细胞白血病细胞中丙酮酸脱氢酶激酶1(pyruvate dehydrogenase kinase 1,PDK1)的mRNA水平。采用基因沉默构建siRNA HIF-1α质粒,分别转染作用于U937和U937/DNR细胞24 h;噻唑蓝(MTT)比色法检测细胞增殖抑制,qPCR检测PDK1 mRNA表达量,蛋白印迹法(Western blotting)检测PDK1和多药耐药基因1(multi-drug resistance gene 1,MDR1)蛋白的表达,流式细胞术检测染料JC-1水平以评估线粒体膜电位变化,血气分析仪测定培养液中乳酸含量。用二氯乙酸盐(dichloroacetate,DCA)和柔红霉素(daunorubicin,DNR)作用于U937/DNR和原代急性单核细胞白血病细胞24 h;MTT比色法检测细胞增殖抑制,Western blotting检测PDK1和MDR1蛋白的表达。结果·PDK1 mRNA在原代急性单核细胞白血病细胞、U937细胞和U937/DNR细胞中均高表达。沉默低氧诱导因子-1α(hypoxiainducible factor-1α,HIF-1α)可显著抑制U937和U937/DNR细胞的增殖活力、PDK1和MDR1蛋白的表达以及乳酸的生成。DCA能够逆转U937/DNR细胞和已复发的原代急性单核细胞白血病细胞对DNR的耐药。结论·HIF-1α-PDK1信号系统可调控细胞糖代谢并参与急性单核细胞白血病的耐药。Objective · To investigate the mechanism of HIF-1α-PDK1 signaling system mediated glucose metabolism and drug resistance in acute monocytic leukemia cells. Methods · The expression of pyruvate dehydrogenase kinase 1 （PDK1） mRNA in U937, U937/DNR and acute monocytic leukemia cells was detected by quantitative polymerase chain reaction （qPCR）. siRNA HIF-1α plasmid was constructed and transferred to U937 and U937/DNR cells for 24 h by gene silencing. Cell proliferation inhibition was examined by MTT assay. The level of PDK1 mRNA was detected by qPCR, and the expression of PDK1 and multi-drug resistance gene 1 （MDR1） proteins was detected by Western blotting. Cell membrane potential was measured by fow cytometry using JC-1. Lactic acid level in the culture fuid was determined by blood gas analyzer. Dichloroacetate （DCA） and daunorubicin （DNR） were added to treat U937/DNR and acute monocytic leukemia cells for 24 h, MTT was used to calculate cell proliferation inhibition and Western blotting was used to estimate the expression of PDK1 and MDR1 proteins. Results · PDK1 mRNA was highly expressed in U937, U937/DNR and acute monocytic leukemia cells. Silencing hypoxia-inducible factor-1α （HIF-1α） signifcantly inhibited the proliferation activity, PDK1 and MDR1 expression and lactic acid production in U937 and U937/DNR cells. DCA could reverse the resistance to DNR in U937/DNR and relapsed acute monocytic leukemia cells. Conclusion · HIF-1α-PDK1 signaling system may regulate glucose metabolism and participate in the drug resistance of acute monocytic leukemia.

关 键 词：丙酮酸脱氢酶激酶1 多药耐药 低氧诱导因子-1Α 急性单核细胞白血病 

分 类 号：R557.2[医药卫生—血液循环系统疾病]