IL-7介导的JAK/STAT信号通路在白藜芦醇抗急性T淋巴细胞白血病中的作用研究

The Mechanism of Resveratrol on Acute T-Lymphocyte Leukemia through IL-7-Mediated JAK/STAT Signaling Pathway

目的:观察T-ALL细胞株和小鼠模型中IL-7及JAK/STAT通路中信号分子的变化,探讨白藜芦醇抗T-ALL的作用机制。方法:将T-ALL细胞株Molt4分成对照、DMSO处理和白藜芦醇处理(Res)共3组,其中对照组不做任何处理,DMSO组用0.05%DMSO处理48 h,Res组用200μmol/L白藜芦醇处理48 h。将15只6-8周龄健康雌性C57BL/6J小鼠随机分为正常对照、T-ALL模型(T-ALL)和白藜芦醇处理(Res)共3组,其中对照和T-ALL组均以0.05%DMSO灌胃处理,Res组以10 mg/ml白藜芦醇灌胃处理。采用RT-qPCR法检测各组细胞和小鼠脾脏组织中IL-7、IL-7R及靶基因Pim1 mRNA的表达,CCK-8法检测细胞的增殖情况,Western blot检测各组细胞和小鼠脾脏组织JAK1、JAK3、STAT5、Pim1及磷酸化JAK1(p-JAK1)、磷酸化JAK3(p-JAK3)、磷酸化STAT5(p-STAT5)的蛋白表达,ELISA法检测各组细胞和小鼠血清中IL-7、IL-7R水平。结果:Res抑制Molt4细胞增殖,下调Molt4细胞及T-ALL小鼠脾脏中p-JAK1、p-JAK3、p-STAT5、Pim1蛋白相对表达量以及Pim1、IL-7、IL-7R mRNA的表达水平,降低Molt4细胞及T-ALL小鼠血清中IL-7、IL-7R的水平(P<0.05)。结论:白藜芦醇可能通过下调IL-7和IL-7R的表达水平,抑制其介导的JAK/STAT信号通路,进而降低靶蛋白Pim1表达发挥抗T-ALL的作用。

Objective:To explore the roles and relative mechanism of resveratrol against T-ALL through detecting the signaling molecules in IL-7 and JAK/STAT pathway.Methods:In vitro experiments,Molt4 cells were divided into 3 groups,including the control group,the DMSO group and resveratrol-treated group(Res group).The control group cells without any treatment,the DMSO group cells treated with 0.05%DMSO for 48 hours,the Res group cells treated with 200μmol/L resveratrol for 48 hours.In vivo experiments,female C57 BL/6 J mice(6-8 weeks)were randomly divided into the control group,the T-ALL model group(T-ALL group),and Res treatment group(Res group).The control group mice treated with 0.05%DMSO by intragastric treatment,the T-ALL group mice treated with 0.05%DMSO by intragastric treatment,and the Res group mice treated with 10 mg/ml resveratrol.Expression of IL-7,IL-7 R and Pim1 mRNA in the cells and mice spleen tissues were detected by RT-qPCR.Cell proliferation ability was detected by CCK-8.The expression of JAK1,JAK3,STAT5,phosphorylated JAK1(p-JAK1),phosphorylated JAK3(p-JAK3),phosphorylated STAT5(p-STAT5)and Pim1 were detected by Western blot.ELISA was used to detect the IL-7 and IL-7 R in the cells and mice serum of each groups.Results:Resveratrol could inhibit the proliferation ability of Molt4 cells,decrease the relative levels of p-JAK1,p-JAK3,p-STAT5,Pim1 protein,and the expression levels of Pim1,IL-7 and IL-7 R mRNA in cells and mice spleens,reduce the IL-7 and IL-7 R in Molt4 cells and mice serum.Conclusion:Resveratrol may inhibite IL-7-medicated JAK/STAT signaling pathway to reduce the expression of target protein Pim1 to further exert its anti-T-ALL effects.