[Effect of arsenic trioxide and 5-aza-2'-deoxycytidine on SHP-1, JAK3, TYK2 gene expression in K562 cells].
This study was purposed to explore the effects of a methylation inhibitor arsenic trioxide (As2O3, 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 pathogenesis 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 1 µmol/L + 5-aza-CdR 0.5 µmol/L, ATO 2.5 µmol/L + 5-aza-CdR 1 µmol/L, and ATO 5 µmol/L + 5-aza-CdR 2 µmol/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 decreased 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-regulated and the expressions of JAK3, TYK2 are down-regulated in concentration-and time-dependent manners, moreover the ATO and 5-aza-CdR show synergies demethylation effect. The SHP-1 gene exert effect possibly through inhibiting the JAK/STAT pathway, the JAK3 is affected more than TYK2, the JAK3 may exert more important role in TAK/STAT pathway.
Department of Hematology, Xingtai people's Hospital of Hebei Medical University , Xingtai, 054000, Hebei Province, China. E-mail: farflybird@163.com.
Xiao-Kun
XK
Zhang
Hebei Province Key Laboratory of Blood Disease, Department of Hematology, The Second Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei Province, China.
Jian-Min
JM
Luo
Department of Hematology, Xingtai people's Hospital of Hebei Medical University , Xingtai, 054000, Hebei Province, China.
Jie
J
Sun
D000077237:Arsenic Trioxide; D001152:Arsenicals / Q000494:pharmacology*; D001374:Azacitidine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D019175:DNA Methylation; D000077209:Decitabine; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D006801:Humans; D053616:Janus Kinase 3 / Q000378:metabolism*; D020014:K562 Cells; D010087:Oxides / Q000494:pharmacology*; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6 / Q000378:metabolism*; D053634:TYK2 Kinase / Q000378:metabolism*
D001152:Arsenicals; D010087:Oxides; D000077209:Decitabine; C507933:JAK3 protein, human; D053616:Janus Kinase 3; D053634:TYK2 Kinase; C070139:TYK2 protein, human; C509249:PTPN6 protein, human; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D001374:Azacitidine; D000077237:Arsenic Trioxide