急性髓系白血病患儿柔红霉素耐药与PD-L1蛋白表达相关

Association between daunorubicin resistanceand PD-L1 protein expression in children with acute myeloid leukemia

目的研究急性髓系白血病(AML)患儿柔红霉素(DNR)耐药与程序性死亡受体配体-1(PD-L1)蛋白表达的相关性。方法选取2016年1月至2022年12月郑州大学附属儿童医院收治的110例AML患儿骨髓样本作为研究组,以50名骨髓正常供体骨髓样本作为对照组。培养人AML细胞系HL60、THP-1、U-937、Molm-13,Western blot检测PD-L1蛋白表达量。构建LV-PD-L1-shRNA、LV-PD-L1-WT-OE慢病毒载体,分析PD-L1对Molm-13细胞DNR耐药的影响及机制。结果研究组PD-L1蛋白表达量高于对照组,AML细胞系中PD-L1蛋白表达量高于健康骨髓单个核细胞(BMMC)(P<0.05),PD-L1表达与AML患儿白细胞计数、骨髓原始细胞比率、预后危险分层、两个标准化学治疗方案后疾病缓解情况有关(P<0.05)。PD-L1高表达组总生存率低于PD-L1低表达组(P<0.05)。与LV-PD-L1-WT-OE组比较,LV-PD-L1-shRNA组PD-L1 mRNA表达量降低、细胞增殖活性降低、凋亡率升高(P<0.05),LV-PD-L1-shRNA可提高Molm-13细胞对DNR的敏感性。TCGA数据库分析显示,6-磷酸葡萄糖脱氢酶(G6PD)可能为PD-L1潜在的目标基因。结论PD-L1在儿童AML中高表达,与患儿化疗耐药有关,其可能通过调控G6PD引起DNR耐药。

Objective To explore the correlation between daunorubicin(DNR)resistance and expression of programmed death receptor ligand 1(PD-L1)in children with acute myeloid leukemia(AML).Methods Totally 110 bone marrow samples were selected from AML patients admitted to Zhengzhou University Affiliated Children′s Hospital from January 2016 to December 2022 as the study group,50 bone marrow samples from normal bone marrow donors were used as the control group.Human AML cell lines HL60,THP-1,U-937,and Molm-13 were cultured,and the expression of PD-L1 protein was detected by Western blot.LV-PD-L1-shRNA and LV-PD-L1-WT-OE lentiviral vectors were constructed.The effect and mechanism of PD-L1 on DNR resistance in Molm-13 cells were analyzed.Results The expression level of PD-L1 protein was higher than that in control group and the expression level of PD-L1 protein in AMI cell lines was higher than that in healthy bone marrow mononuclear cells(BMMC)(P<0.05).PD-L1 expression was related to white blood cell count,bone marrow primitive cell ratio,prognostic risk stratification and disease remission after two standard chemotherapy regimens in AML patients(P<0.05).The overall survival rate of the PD-L1 high expression group was lower than that of the PD-L1 low expression group(P<0.05).Compared with the LV-PD-L1-WT-OE group,the LV-PD-L1-shRNA group showed a decrease in PD-L1 mRNA expression,cell proliferation activity but an increased apoptosis rate(P<0.05).LV-PD-L1-shRNA enhanced sensitivity of Molm-13 cells to DNR.TCGA database analysis showed that glucose 6-phosphate dehydrogenase(G6PD)was a potential target gene for PD-L1.Conclusions The high expression of PD-L1 in pediatric AML is related to chemotherapy resistance in children with AML,and DNR resistance might be caused through regulation of G6PD.