三氧化二砷联合5-杂氮脱氧胞苷对K562细胞SHP-1、JAK3、TYK2基因表达的影响  

Effect of Arsenic Trioxide and 5-Aza-2'-deoxycytidine on SHP-1,JAK3,TYK2 Gene Expression in K562 Cells

作　　者：张小坤[1] 罗建民[2] 孙杰[1] 

机构地区：[1]河北医科大学附属邢台人民医院血液科,河北邢台054000 [2]河北医科大学第二医院血液科、血液病河北省重点实验室,河北石家庄050011

出　　处：《中国实验血液学杂志》2014年第2期323-328,共6页Journal of Experimental Hematology

摘　　要：本研究旨在探讨甲基化抑制剂三氧化二砷(As2O3)及5-杂氮脱氧胞苷(5-aza-CdR)对JAK-STAT信号转导通路中JAK家族成员JAK3、TYK2和造血细胞磷酸酶SHP-1在慢性髓系白血病细胞株K562中表达的影响及它们在白血病发病中的作用。K562细胞分为单药组、联合用药组及不加药物组(空白对照)。5-aza-CdR单用浓度为0.5、1、2μmol/L,As2O3单用浓度为1、2.5、5μmol/L,As2O31μmol/L+5-aza-CdR 0.5μmol/L、As2O32.5μmol/L+5-aza-CdR 1μmol/L。As2O3联合用药浓度为5μmol/L+5-aza-CdR 2μmol/L。对细胞分别处理24、48、72 h后提取细胞总RNA,应用荧光实时定量PCR(Real-time quantitative polymerase chain reaction,RQ-PCR)检测JAK3、TYK2及SHP-1的表达。结果表明,As2O3和5-aza-CdR单独作用及两药合用时,SHP-1 mRNA在K562细胞中的表达呈剂量及时间依赖性升高,JAK3 mRNA的表达呈剂量及时间依赖性降低,TYK2 mRNA的表达呈剂量及时间依赖性降低,SHP-1与JAK3、TYK2呈负相关,JAK3所受影响较TYK2更为显著。结论:As2O3和5-aza-CdR单独及联合应用时,随着浓度增高和作用时间延长,SHP-1表达上调,JAK3和TYK2表达下调,且两药有协同去甲基化作用;SHP-1基因可能通过抑制JAK/STAT通路发挥作用,JAK3所受影响较TYK2更为显著。在JAK/STAT通路中,JAK3可能发挥更为重要的作用。This study was purposed to explore the effects of a methylation inhibitor arsenic trioxide （As203 ,ATO） and 5-Aza-2＇-deoxycytidine （5-aza-CdR） on the expression of JAK-STAT signal transduction pathway in family members JAK3, TYK2 and hematopoietic cell phosphatase SHP-1 in chronic myeloid leukemia cell line K562 and their roles in pathgenesis of leukemia. The K562 cells were divided into 3 groups： single drug-treated group, combined 2 drugs-treated group, group without drug treatment as control. The concentration of 5-aza-CdR were 0.5, 1,2 μmol/L; the concentration of ATO was 1, 2.5, 5 μmol/L; the concentration of combined drugs was ATO 1μmol/L ＋ 5-aza- CdR 0.5 μmol/L, ATO 2.5μ mol/L ＋ 5-aza-CdR 1μmol/L, and ATO 5 μmol/L ＋ 5-aza-CdR 2μmol/L. The K562 cells were treated with above-mentioned concentration of drugs for 24, 48 and 72 hours, then the total RNA of cells was extracted, the JAK3, TYK2 and SHP-1 expressions were detected by real-time quantitative-PCR. The results showed that after the K562 cells were treated with ATO and 5-aza-CdR alone and their combination, the expression of SHP-1 mRNA increased, the expressions of JAK3 mRNA and TYK2 mRNA decreated along with increasing of concentration and prolonging of time, displaying the concentration and time-dependency. The SHP-1 negatively related with JAK3 and TYK2. The effect of SHP-1 on JAK3 was significantly higher than that on TYK2. It is concluded that when the K562 cells are treated with ATO and 5-aza-CdR alone and their combination, the expression of SHP-1 is up- regalated and the expressions of JAK3, TYK2 are down-regalated in concentration-and time-dependent manners, moreover the ATO and 5-aza-CdR show synergical demethylation effect. The SHP-1 gene exert effect possibly throughinhibiting the JAK/STAT pathway, the JAK3 is affected more than TYK2, the JAK3 may exert more impartant role in TAK/STAT pathway.

关 键 词：甲基化抑制剂 5-杂氮脱氧胞苷 三氧化二砷 K562细胞 SHP-1 JAK3 TYK2 白血病 

分 类 号：R733.7[医药卫生—肿瘤] R979.1[医药卫生—临床医学]