急性淋巴细胞白血病患儿骨髓单个核细胞中miR-124表达及其功能研究
Expression of miR-124 in bone marrow mononuclear cells of children with acute lymphoblastic leukemia and its function
褚静 1张楠 2李晓云 2胡勇3
作者信息
- 1. 潍坊市益都中心医院儿童康复科,山东潍坊 262500
- 2. 潍坊市益都中心医院神经康复科
- 3. 山东第二医科大学口腔学院
- 折叠
摘要
目的 探讨急性淋巴细胞白血病(ALL)患儿骨髓单个核细胞中miR-124表达及其在CCRF-CEM和CEM-C1细胞株中的作用.方法 选取ALL患儿106例(初诊患儿65例、完全缓解患儿26例和难治复发患儿15例)及行骨髓穿刺检查且骨髓像正常的儿童25例作为对照组,RT-PCR技术检测骨髓单个核细胞中miR-124表达;用电穿孔转染法对CCRF-CEM细胞转染miR-124抑制物和阴性对照物,对CEM-C1细胞转染miR-124模拟物和阴性对照序列,RT-PCR技术检测各组细胞miR-124表达;分别给予不同终浓度的喜树碱处理,CCK-8法检测各组细胞增殖情况,计算增殖抑制率和细胞半数抑制浓度(IC50).结果 与对照组相比,ALL患儿的骨髓单个核细胞中miR-124相对表达量均降低(P＜0.05),且初诊患儿miR-124相对表达量低于完全缓解患儿,高于难治复发患儿(P＜0.05);初诊患儿的骨髓单个核细胞中miR-124表达量与WBC计数、危险度分层和脾肿大有关(P＜0.05);转染miR-124抑制物的CCRF-CEM细胞IC50=774.17 μg/L,转染miRNA抑制物阴性对照的细胞IC50=27.82 μg/L,RI=27.83;转染miR-124模拟物的CEM-C1细胞IC50=1.06 μg/L,转染阴性对照序列的细胞IC50=199.52 μg/L,RI=188.23.结论 miR-124在ALL患儿骨髓单个核细胞中表达降低,可能参与了 ALL患儿复发及耐药过程.
Abstract
Objective To investigate the expression of miR-124 in bone marrow mononuclear cells of chil-dren with acute lymphoblastic leukemia(ALL),and explore its function in CCRF-CEM and CEM-C1 cell lines.Methods One hundred and six cases of children with ALL were selected(including 65 cases of newly diagnosed,26 cases of complete remission and 15 cases of refractory relapsed),and 25 cases who underwent bone marrow puncture examination and had normal bone marrow images were selected as the control group.The expressions of miR-124 in bone marrow mononuclear cells of children with ALL and control group were detected by RT-PCR.The CCRF-CEM cells were trans-fected with miR-124 inhibitor and miRNA inhibitor negative control by electroporation,and CEM-C1 cells were transfect-ed with miR-124 mimics and negative control sequences.The expression of miR-124 in cells of each group was detected by RT-PCR.After giving different final concentrations of camptothecin treatment,CCK-8 method was used to detect the cell proliferation in each group,and the cell proliferation inhibitory rate and half-cell inhibitory concentration(IC50)were calculated.Results Compared with the control group,the relative expression levels of miR-124 in the bone marrow mononuclear cells of children with newly diagnosed,complete remission and refractory relapsed were decreased(P＜0.05).The relative expression level of miR-124 in bone marrow mononuclear cells of children with newly diagnosed was lower than that in the children with complete remission,while was higher than that in the children with refractory re-lapsed,the differences were statistically significant(P＜0.05).The expression of miR-124 in bone marrow mononuclear cells of children with newly diagnosed was associated with WBC count,risk stratification and splenomegaly(P＜0.05).IC50 in CCRF-CEM cells transfected with miR-124 inhibitor was 774.17 μg/L,and in cells transfected with miRNA in-hibitor negative control was 27.82 μg/L,and RI was 27.83.IC50 in CEM-C1 cells transfected with miR-124 mimics was 1.06 μg/L,and in cells transfected with negative control sequences was 199.52 μg/L,and RI was 188.23.Conclusion The expression of miR-124 in bone marrow mononuclear cells of children with ALL was decreased.It might be in-volved in relapse and drug resistance process of ALL children.
关键词
儿童/淋巴细胞白血病,急性/miR-124/骨髓单个核细胞/耐药性Key words
Child/Acute lymphoblastic leukemia/Mir-124/Bone marrow mononuclear cells/Drug resistance引用本文复制引用
出版年
2024
NETL
NSTL
万方数据