MiR-155对小儿急性淋巴细胞白血病的耐药机制研究  

Mechanism of miR-155 Promoting Drug Resistance in Childhood Acute Lymphoblastic Leukemia by Regulating Wnt/β-Catenin Signaling Pathway

作　　者：黄惠敏 韦云剑 王丹[1] 温晓梅[1] HUANG Hui-Min;WEI Yun-Jian;WANG Dan;WEN Xiao-Mei(Depariment of Pediatrics,The First Affiliated Hospital of Hainan Medical University,Haikou 570100,Hainan Province,China;Hainan Province Key Laboratory For Human Reproductive Medicine And Genetic Research,Haikou 570100,Hainan Province,China)

机构地区：[1]海南医学院第一附属医院儿科,海南海口570100 [2]海南医学院第一附属医院海南省人类生殖与遗传重点实验室,海南海口570100

出　　处：《中国实验血液学杂志》2022年第2期418-424,共7页Journal of Experimental Hematology

摘　　要：目的:探讨miR-155通过调节Wnt/β-Catenin信号通路促进小儿急性B淋巴细胞白血病(B-ALL)对阿糖胞苷(Ara-C)化疗耐药的机制。方法:采用PCR检测B-ALL患儿骨髓组织与细胞系中miR-155表达情况;用REH细胞系构建化疗抵抗株REH/Ara-C,使用miR-155抑制剂转染REH/Ara-C细胞,EdU检测细胞增殖能力,流式细胞术检测细胞的凋亡率,CCK-8法与克隆形成实验分析细胞耐药性,Western blot测定Wnt/β-Catenin信号通路相关蛋白的表达;用miR-155抑制剂与Wnt激活剂agonist转染REH/Ara-C细胞,测定其对细胞增殖、凋亡与耐药性的影响。结果:与正常组织及细胞相比,B-ALL骨髓组织/细胞系中miR-155表达水平升高(P<0.05);与药物敏感B-ALL组织/细胞系相比,耐药B-ALL组织与细胞系中miR-155表达水平升高(P<0.05);抑制miR-155表达降低了REH/Ara-C细胞增殖能力(P<0.05),促进了凋亡(P<0.05),增强了Ara-C的细胞毒性(P<0.05),同时抑制了Wnt/β-Catenin信号通路相关蛋白与耐药基因MDR1的表达(P<0.05),通过激活Wnt表达则可逆转上述现象(P<0.05)。结论:B-ALL患儿骨髓组织中miR-155表达上调,可以通过激活Wnt/β-Catenin信号通路促进B-ALL细胞增殖、抑制细胞凋亡,从而导致化疗耐药。Objective: To investigate the mechanism of miR-155 promoting drug resistance of children B-ALL to Ara-C by regulating Wnt/β-Catenin signaling pathway. Methods: The expression of miR-155 in bone marrow tissue and cell line of B-ALL was detected by PCR. The chemotherapy resistant strain REH/Ara-C was constructed by using REH cells. REH/Ara-C cells were transfected with miR-155 inhibitor. The proliferation of REH/Ara-C cells was detected by EdU. The apoptosis of REH/Ara-C cells was detected by flow cytometry. The drug resistance of REH/Ara-C cells were analyzed by CCK-8 method and colony formation assay. The expression of Wnt/β-Catenin signaling pathway related proteins were determined by Western blot. MiR-155 inhibitor and Wnt activator agonist were used to transfect REH/Ara-C cells, and their effects on cell proliferation, apoptosis and drug resistance were determined. Results: Compared with normal tissues and cells, the expression level of miR-155 in B-ALL bone marrow tissue/cell line was increased(P<0.05);Compared with drug sensitive B-ALL tissues/cell lines, the expression level of miR-155 in drug resistant B-ALL tissues and cell lines was increased(P<0.05);Inhibition of miR-155 expression decreased the proliferation of REH/Ara-C cells(P<0.05), promoted apoptosis(P<0.05), enhanced the cytotoxicity of Ara-C(P<0.05), and inhibited Wnt/β-Catenin signaling pathway related protein and MDR1 gene expression(P<0.05), which could be reversed by activating Wnt expression(P<0.05). Conclusion: The expression of miR-155 is up-regulated in bone marrow of children with B-ALL, which may be related to the activation of Wnt/β-Catenin signaling pathway promotes the proliferation of B-ALL cells and inhibits apoptosis, which leads to chemotherapy resistance.

关 键 词：MIR-155 WNT/Β-CATENIN 急性B淋巴细胞白血病 小儿 化疗耐药 

分 类 号：R733.71[医药卫生—肿瘤]