PPARD通过JAK2/STAT3信号通路对急性髓系白血病细胞增殖的作用

Study on the effect and mechanism of PPARD on acute myeloid leukemia through JAK2/STAT3 signal pathway

目的探讨PPARD基因在急性髓系白血病(AML)患者骨髓中的表达和意义并分析PPARD与JAK-STAT信号通路的关系及对AML细胞株THP-1增殖的影响。方法收集我院初诊AML患者和非血液疾病患者的临床资料和骨髓标本,采用RT-qPCR、Western blot分别检测AML患者中PPARD mRNA和蛋白表达,并分析其表达与患者性别、年龄、骨髓幼稚细胞百分比、白细胞数等的关系;培养THP-1细胞至对数生长期,用不同浓度(0、0.1、1、5、10μM)PPARD抑制剂(GSK3787)分别作用THP-1细胞24、48、72 h后,CCK-8法检测THP-1细胞的增殖,确定最佳药物作用浓度及时间;激活及抑制PPARD后分别检测JAK2、STAT3的表达,分析PPARD与JAK2/STAT3信号通路的关系。结果收集AML患者46例,对照组17例,AML患者PPARD在mRNA表达水平上低于正常对照组,差异有统计学意义(P<0.05),且与AML患者的年龄、性别和FBA分型有关,而在蛋白水平上高于正常对照组,差异有统计学意义(P<0.05);抑制PPARD后,JAK2、STAT3表达均明显下降,激活PPARD表达后,JAK2、STAT3蛋白表达均增加,均具有统计学差异(P<0.05)。结论AML患者中PPARD mRNA低于对照组,蛋白高于对照组;PPARD可能通过调控JAK2/STAT3信号通路的表达来影响THP-1细胞的增殖。

Objective To investigate the expression and significance of PPARD gene in the bone marrow of patients with acute myeloid leukemia(AML)and to analyze the relationships of PPARD with JAK-STAT signaling pathway and its impacts on the proliferation of AML cell line THP-1 cells.Methods A total of newly diagnosed with AML and non-hematoligical diseases were selected.The clinical data and bone marrow specimens were collected.The expression of PPARD mRNA and protein by RT-qPCR and Western blot,respectively.The correlations between the expression of PPARD and the clinical characteristics were analyzed.THP-1 was treated with PPARD inhibitor(GSK3787)at concentrations of 0,0.1,1,5,10μM for 24,48,72 h,respectively.The changes of cell proliferation were detected by CCK8 for the determination of best drug concentration and time.After activating and inhibiting PPARD,the expressions of JAK2 and STAT3 were detected,and the relationship between PPARD and JAK2/STAT3 signaling pathway was analyzed.Results A total of AML patients 46 cases and control group 17 cases were collected,The expression level of PPARD mRNA in AML patients was lower than that of control group(P<0.05),which was related to the age,gender and FBA classification of AML patients(P<0.05).The expression level of PPARD protein in AML patients was higher than that of the control group(P<0.05).After inhibiting PPARD,the expression of JAK2 and STAT3 decreased significantly(P<0.05).After activating the expression of PPARD,the expression of JAK2 and STAT3 protein increased(P<0.05).Conclusion In AML patients,PPARD mRNA is lower than the control group,but the protein level is higher than the control group.PPARD may affect the proliferation of THP-1 cells by regulating the expression of JAK2/STAT3 signaling pathway.