| 侯秋苹,任妍,付敏,等.微小 RNA?34a调控儿童急性淋巴细胞白血病细胞 CEM/C1生长和迁移的分子机制研究[J].安徽医药,2020,24(12):2512-2515. |
| 微小 RNA?34a调控儿童急性淋巴细胞白血病细胞 CEM/C1生长和迁移的分子机制研究 |
| Study on the molecular mechanism of miR?34a regulating the growth and migration of CEM/C1 cells in acute lymphoblastic leukemia |
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| DOI：10.3969/j.issn.1009?6469.2020.12.045 |
| 中文关键词: 白血病 转染 细胞增殖 急性淋巴细胞白血病 人急性淋巴细胞白血病细胞 CEM/C1 微小 RNA?34a Krüppel样因子 4 儿童 |
| 英文关键词: Leukemia Transfection Cell proliferation Acute lymphoblastic leukemia Human acute lymphoblastic leuke? mia cells CEM/C1 microRNA?34a Krüppel?like factor 4 Child |
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| 中文摘要: |
| 目的研究微小 RNA?34a(miR?34a)对儿童急性淋巴细胞白血病(ALL)细胞 CEM/C1增殖、迁移的影响以及可能作用机制。方法选取重庆市第十三人民医院血液科 2017年 10月至 2018年 12月住院 32例 ALL病儿骨髓样本。另选取该院同期 25例原发性血小板减少症病儿的骨髓样本作为对照组。实时荧光定量多聚核苷酸链式反应(qPCR)检测 miR?34a在 ALL骨髓样本的表达量； CEM/C1细胞按随机数字表法分为对照组、阴性对照(NC)组、 miR?34a组；对照组细胞常规培养， NC组和 miR?34a组 CEM/C1细胞分别在 Lipofectamine 2000介导下转染阴性对照质粒以及 miR?34a模拟物， qPCR检测转染效率；细胞计数试剂盒 8(CCK?8)实验检测 CEM/C1细胞的增殖， Transwell检测细胞迁移；在线软件 TargetScan预测 miR?34a与 Krüppel样因子 4(KLF4)的靶向关系，双荧光素酶报告基因进一步进行验证。结果与对照组相比， ALL骨髓样本中 miR?34a的表达量下调［(0.33±0.05)比(1.00±0.08)t＝15.680，P＜0.001］。与对照组(1.00±0.08)相比， NC组 miR?34a的表达量(0.99±0.08)无明显变化， miR?34a组 CEM/C1细胞中，miR?34a的表达量(3.21±0.23)上调(F＝423.135，P＜0.05)。对照组、 NC组、 miR?34a组细胞培养 48 h后细胞活性分别为(0.65±0.05)、(0.69±0.06)、(0.42±0.04)F＝40.818，P＜0.001；迁移细胞数分别为(142.36±12.47)个、(137.45±13.49)个、(79.89±6.23)个， F＝57.683，P＜0.001，差异计学意义。与 NC与 KLF4?wt共转染的细胞相比， miR?34a有统，与 KLF4?wt共转染的细胞荧光素酶活性［(0.45±0.03)比(1.00±0.06)t＝20.083，P＜0.001］降低；与 NC与 KLF4?mut共转染的细胞相比， miR?34a与 KLF4?mut共转染的细胞荧光素酶活性差异无统计学，意义［(1.03±0.07)比(1.01±0.07)，t＝0.495，P＞0.05)］。结论 miR?34a在 ALL中表达量下调，其可通过对靶基因 KLF4的调控影响 CEM/C1细胞增殖、迁移。 |
| 英文摘要: |
| Objective To study the effects of microRNA?34a(miR?34a)on the proliferation and migration of CEM/C1 cells in acute lymphoblastic leukemia(ALL)and its possible mechanism.Methods The expression of miR?34a in ALL cases were detect? ed by quantitative polymerase chain reaction(qPCR).CEM/C1 cells were randomly assigned into control group，negative control(NC)group and miR?34a group.The cells of control group were cultured routinely.CEM/C1 cells in NC group and miR?34a groupwere transfected into negative control and miR?34a mimics by Lipofectamine 2000，respectively.The transfection efficiency was de? tected by qPCR.The cell proliferation was observed by cell counting kit?8(CCK?8)and the migration was analyzed by Transwell. TargetScan，an online software，predicted the targeting relationship between miR?34a and Krüppel?like factor 4(KLF4)，and further validates by dual luciferase reporter gene.Results Compared with the control group，the expression of miR?34a in ALL bone mar? row samples was down?regulated［(0.33±0.05)vs.(1.00±0.08)t＝15.680，P＜0.001］.Compared with the control group，the expres? sion of miR?34a in the NC group did not change significantly［(，1.00±0.08)vs.(0.99±0.08)］，and the expression of miR?34a in CEM/C1 cells in the miR?34a group was up?regulated［(1.00±0.08)vs.(3.21±0.23)，F＝423.135，P＜0.05］.The cell viability of the control group，NC group and miR?34a group after 48 hours of cell culture were(0.65±0.05)，(0.69±0.06)，and(0.42±0.04)，re? spectively(F＝40.818，P＜0.001)； the number of migrating cells were(142.36±12.47)(137.45±13.49)and(79.89±6.23)，re? spectively(F＝57.683，P＜0.001)，thedifferencewasstatisticallysignificant.Comparedwit，hthecellsco?tran，sfected with NC and KLF4?wt，the luciferase activity of cells co?transfected with miR?34a and KLF4?wtwas decreased［(0.45±0.03)vs.(1.00±0.06)，t＝20.083，P＜0.001］，the difference was statistically significant.Compared with the cells co?transfected with NC and KLF4?mut，there was no statistically significant difference in luciferase activity in the cells co?transfected with miR?34a and KLF4?mut［(1.03± |
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