miR-133a-3p靶向TGIF2调节急性髓系白血病细胞系OCI-AML3细胞凋亡与线粒体功能

doi:10.3870/j.issn.1672-8009.2026.04.001

摘要/Abstract

摘要： 目的分析miR-133a-3p及其潜在靶点转化生长因子-β诱导因子2(transcription growth factor-β-induced factor 2,TGIF2)在急性髓系白血病细胞系OCI-AML3细胞增殖、凋亡及线粒体功能中的作用机制。方法 OCI-AML3细胞分别或共转染miR-133a-3p mimic和pcDNA-TGIF2。RT-qPCR检测转染效率;生物信息学软件预测miR-133a-3p和TGIF2靶向位点;利用双荧光素酶报告系统确认二者靶向关系;CCK-8实验和EdU染色检测细胞增殖;流式细胞术分析细胞凋亡水平与线粒体膜电位;试剂盒检测ROS含量;蛋白质印迹检测Caspase-3、Caspase-9、TGIF2、Bcl-2、Bax及c-Myc蛋白的表达。结果在OCI-AML3细胞中,miR-133a-3p与TGIF2的表达呈负相关,miR-133a-3p靶向结合TGIF2。与对照组比较,TGIF2组的细胞活力显著升高(P<0.05);EdU阳性细胞明显增多;细胞凋亡率显著降低(P<0.05);Bax/Bcl-2、Caspase-3及Caspase-9表达显著下调(P<0.05),而c-Myc表达显著上调(P<0.05);线粒体膜电位无明显变化;ROS含量显著降低(P<0.05)。miR-133a-3p过表达逆转了上述结果。与TGIF2组比较,mimic+TGIF2组的细胞活力显著降低(P<0.05);EdU阳性细胞明显减少;细胞凋亡率显著升高(P<0.05);Bax/Bcl-2、Caspase-3及Caspase-9表达显著上调(P<0.05),而c-Myc表达显著下调(P<0.05);线粒体膜电位明显改变;ROS含量显著升高(P<0.05)。结论 miR-133a-3p通过靶向TGIF2抑制OCI-AML3细胞活力及增殖,上调ROS水平,诱导线粒体膜电位去极化,促进细胞凋亡。

关键词: miR-133a-3p, 转化生长因子-β诱导因子2, 急性髓系白血病, 细胞凋亡, 活性氧

Abstract: Objective To analyze the effect of miR-133a-3p and its potential target transforming growth factor-β-induced factor 2(TGIF2)on the apoptosis and mitochondrial function of acute myeloid leukemia cells OCI-AML3. Methods OCI-AML3 cells were transfected with miR-133a-3p mimic or/and pcDNA-TGIF2.Transfection efficiency was detected by RT-qPCR.The target sites of miR-133a-3p and TGIF2 were predicted by bioinformatics,and the targeting relationship was verified by dual-luciferase reporter gene assay.Cell proliferation was detected by CCK-8 assay and EdU staining.Flow cytometry was applied to detect the apoptosis and mitochondrial membrane potential.ROS level was measured by associated kit.Western blot was used to detect the expression levels of Caspase-3,Caspase-9,TGIF2,Bcl-2,Bax and c-Myc proteins. Results In OCI-AML3 cells,the miR-133a-3p expression was negatively correlated with TGIF2 expression,and miR-133a-3p targetly binded with TGIF2.Compared with those in the control group,the cell viability in the TGIF2 group was significantly enhanced(P<0.05),the number of EdU positive cells was increased significantly,while the apoptosis rate was significantly reduced(P<0.05);the expressions of Bax/Bcl-2,Caspase-3 and Caspase-9 were significantly down-regulated(P<0.05),while the expression level of c-Myc was significantly up-regulated(P<0.05);the mitochondrial membrane potential showed no significant change,whereas ROS content demonstrated a marked decrease(P<0.05).The overexpression of miR-133a-3p reversed the above results.Compared with those in the TGIF2 group,the mimic+TGIF2 group exhibited significantly reduced cell viability(P<0.05),the number of EdU positive cells was significantly declined,and the apoptosis rate was significantly increased(P<0.05);The expression levels of Bax/Bcl-2,Caspase-3 and Caspase-9 were significantly up-regulated(P<0.05),while the expression level of C-MYC was significantly down-regulated(P<0.05);the mitochondrial membrane potential was significantly changed,and ROS level was significantly increased(P<0.05). Conclusion miR-133a-3p inhibits OCI-AML3 cell viability and proliferation,up-regulates ROS level,induces mitochondrial membrane potential depolarization,and promotes apoptosis by targeting TGIF2.

Key words: miR-133a-3p, transforming growth factor-β-induced factor 2, acute myeloid leukemia, apoptosis, reactive oxygen species

中图分类号:

R349.5

郭丽, 祝俊玲, 卫雷, 郭巧伟, 龚艳玲. miR-133a-3p靶向TGIF2调节急性髓系白血病细胞系OCI-AML3细胞凋亡与线粒体功能[J]. 医学分子生物学杂志, 2026, 23(4): 375-381.

GUO Li, ZHU Junling, WEI Lei, GUO Qiaowei, GONG Yanling. miR-133a-3p Regulates Apoptosis and Mitochondrial Function of OCI-AML3 Acute Myeloid Leukemia Cells via Targeting TGIF2[J]. Journal of Medical Molecular Biology, 2026, 23(4): 375-381.

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