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。
Objective To investigate the expression of miR-103a-3p in bone marrow tissue of patients with chronic myeloidleukemia(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 apoptosis 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 inhibition 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).ConclusionThis study shows that miR-103a-3p can inhibit the proliferative capacity of CML K562 cells and promote their apoptosis.
作者
王智超
谢文杰
管洪在
WANG Zhichao;XIE Wenjie;GUAN Hongzai(Department of Laboratory,The Affiliated Hospital of Qingdao University,Qingdao 266071,China)
出处
《青岛大学学报（医学版）》
CAS
2024年第2期252-256,共5页
Journal of Qingdao University(Medical Sciences)
基金
山东省自然科学基金面上项目(ZR2020MH311)。
