Objective: To investigate the effect of miR-22 targeting formin-like protein 2 (FMNL2) on the migration and apoptosis of childhood acute myeloid leukemia (AML) cells.
Method: Peripheral blood samples from 11 children with AML, 10 children with immune thrombocytopenia, human AML cell lines TF-1a, HL-60, THP-1 and human bone marrow stromal cells HS-5 were used as the research objects. UniCel DxH 800 automatic hematology analyzer detected platelet count, hemoglobin, and white blood cell count in peripheral blood samples, and RT-qPCR detected miR-22 expression in peripheral blood samples and AML cells. HL-60 cells were transfected with LipofectamineTM 2000 kit, the experiments were divided into seven groups: blank (no cells transfected), miR-NC, miR-22 mimics, si-NC, si-FMNL2 , miR-22 mimics+OE-NC and miR-22 mimics+OE-FMNL2 . RT-qPCR was used to detect the expression of miR-22 in each group. Transwell was used to detect cell migration. Flow cytometry was used to detect cell apoptosis. Dual-luciferase reporter gene detection experiments verified the targeting relationship between miR-22 and FMNL2 . Western blot was used to detect the expression of FMNL2 protein.
Results: Compared with the control group, the number of leukocytes in the peripheral blood of children with AML was significantly increased (P <0.001), while the concentration of hemoglobin and the number of platelets were significantly decreased P <0.001). The expression level of miR-22 in peripheral blood of children with AML was significantly lower than that in control group (P <0.001). Compared with HS-5 cells, the expression levels of miR-22 in TF-1a, HL-60, and THP-1 cells were significantly decreased (P <0.05), and in HL-60 cells was the lowest. Therefore, HL-60 cells were selected for subsequent experiments. Up-regulation of miR-22 or silencing of FMNL2 could reduce the number of migrating cells and increase apoptosis rate (P <0.05). MiR-22 targeted and negatively regulated the expression of FMNL2 . FMNL2 overexpression reversed the effects of up-regulated miR-22 on migration and apoptosis of HL-60 cells.
Conclusion: MiR-22 can inhibit the migration and promote apoptosis of HL-60 cells by down regulating the expression of FMNL2 .
题目: miR-22靶向FMNL2 对儿童急性髓系白血病细胞迁移和凋亡的影响.
目的: 探讨miR-22靶向同源形成素样蛋白2（FMNL2）对儿童急性髓系白血病（AML）细胞迁移和凋亡的 影响。.
方法: 以11例AML患儿、10例免疫性血小板减少患儿的外周血标本以及人AML细胞系TF-1a、HL-60、THP-1和人骨髓基质细胞HS-5为研究对象，UniCel DxH 800型全自动血液分析仪检测外周血标本的血小板数、血红蛋白、白细胞数，RT-qPCR检测外周血标本和AML细胞中miR-22表达；利用LipofectamineTM 2000试剂盒对HL-60细胞进行转染，实验分为空白（细胞未转染）、miR-NC、miR-22 mimics、si-NC、si-FMNL2 、miR-22 mimics+OE-NC 和miR-22 mimics+OE-FMNL2 共7组，RT-qPCR检测各组细胞miR-22表达；Transwell检测细胞迁移；流式细胞术检测细胞凋亡；双荧光素酶报告基因检测实验验证miR-22与FMNL2 靶向关系；Western blot检测FMNL2蛋白表达 情况。.
结果: 与对照组比较，AML患儿外周血中白细胞数显著升高（P <0.001），血红蛋白浓度及血小板数显著降低（P <0.001）；AML患儿外周血中miR-22的表达水平显著低于对照组（P <0.001）；与HS-5细胞比较，TF-1a、HL-60、THP-1细胞中miR-22的表达水平均显著降低（P <0.05），且HL-60细胞中的miR-22的表达水平最低，因此，选择HL-60细胞进行后续实验；上调miR-22或沉默FMNL2 均可降低迁移细胞数目，提高细胞凋亡率（P <0.05）；miR-22靶向负调控FMNL2 的表达；FMNL2 过表达逆转了上调miR-22对HL-60细胞迁移和凋亡的影响。.
结论: miR-22通过靶向下调FMNL2 表达，抑制HL-60细胞迁移、促进细胞凋亡。.
Keywords: acute myeloid leukemia; apoptosis; cell migration; homeomorphin-like protein 2; miR-22.