miR-103a-3p对慢性粒细胞白血病细胞增殖和凋亡影响
Effect of miR-103a-3p on the proliferation and apoptosis of chronic myeloid leukemia cells
王智超 1谢文杰 1管洪在1
作者信息
- 1. 青岛大学附属医院检验科,山东青岛 266071
- 折叠
摘要
目的 检测miR-103a-3p在慢性粒细胞白血病(CML)病人骨髓组织中的表达,探讨miR-103a-3p对CML细胞增殖、凋亡的影响.方法 收集32例CML病人和20例健康供者的骨髓标本,采用实时荧光定量PCR(RT-qPCR)检测miR-103a-3p表达.采用CCK8法和流式细胞术检测CML细胞系K562细胞增殖及凋亡的变化;采用免疫印迹法(Western blot)检测K562细胞中凋亡相关蛋白Bcl-2相关X蛋白(Bax)和B淋巴细胞瘤-2基因(Bcl-2)表达;小鼠皮下注射K562细胞,通过苏木精-伊红(HE)染色观察组织病理变化.结果 与健康供者相比,miR-103a-3p在CML病人的骨髓单个核细胞中的表达量显著降低(t=3.317,P＜0.01);K562细胞中的miR-103a-3p表达水平也显著低于正常人骨髓基质细胞HS-5(t=21.430,P＜0.001).体外实验显示,转染agomiR-103a-3p后,K562细胞的增殖受到抑制(t=4.949～12.170,P＜0.01),凋亡增强(t=3.181,P＜0.05).成瘤实验显示,Lv-miR-up组小鼠的肿瘤质量较Lv-miR-NC组低(t=4.518,P＜0.01),肿瘤体积较Lv-miR-NC组小(t=15.670～36.290,P＜0.001).结论 miR-103a-3p可抑制CML细胞K562的增殖,促进其凋亡.
Abstract
Objective To investigate the expression of miR-103a-3p in bone marrow tissue of patients with chronic my-eloidleukemia(CML)and the effect of miR-103a-3p on the proliferation and apoptosis of CML cells.Methods Bone marrow specimens were collected from 32 patients with CML and 20 healthy donors,and quantitative real-time PCR was used to measure the expression of miR-103a-3p.CCK-8 assay and flow cytometry were used to measure the changes in the proliferation and apopto-sis of CML K562 cells,and Western blot was used to measure the expression levels of the apoptosis-related proteins Bcl2-associated X protein(Bax)and B-cell lymphoma-2(Bcl2)in K562 cells.Mice were subcutaneously injected with K562 cells,and hematoxylin-eosin staining was used to observe histopathological changes.Results Compared with the healthy donors,the patients with CML had a significant reduction in the expression level of miR-103a-3p in bone marrow mononuclear cells(t=3.317,P＜0.01),and the expression level of miR-103a-3p in K562 cells was significantly lower than that in normal human bone marrow stroma cells(t=21.430,P＜0.001).In vitro experiments showed that after transfection with agomiR-103a-3p,K562 cells showed significant inhibi-tion of proliferation(t=4.949-12.170,P＜0.01)and a significant increase in apoptosis(t=3.181,P＜0.05).Tumor formation experiments showed that compared with the Lv-miR-NC group,the Lv-miR-up group had a significantly lower tumor mass(t=4.518,P＜0.01)and a significantly smaller tumor volume(t=15.670-36.290,P＜0.001).Conclusion This study shows that miR-103a-3p can inhibit the proliferative capacity of CML K562 cells and promote their apoptosis.
关键词
微RNAs/白血病,髓系,慢性,BCR-ABL阳性/K562细胞/细胞增殖/细胞凋亡Key words
microRNAs/leukemia,myelogenous,chronic,BCR-ABL positive/K562 cells/cell proliferation/apoptosis引用本文复制引用
基金项目
山东省自然科学基金面上项目(ZR2020MH311)
出版年
2024
国家科技期刊平台
NSTL
万方数据