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微小RNA-34b在小儿急性白血病中的表达的临床意义

Clinical significance of microRNA-34b expression in pediatric acute leukemia.

作者信息

Cao Lan, Wang Na, Pan Jian, Hu Shaoyan, Zhao Wenli, He Hailong, Wang Yi, Gu Guixiong, Chai Yihuan

机构信息

Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, Jiangsu 215003, P.R. China.

出版信息

Mol Med Rep. 2016 Mar;13(3):2777-84. doi: 10.3892/mmr.2016.4876. Epub 2016 Feb 5.

Abstract

The present study aimed to explore the function of miR‑34b promoter methylation in cell proliferation in children's acute leukemia. Quantitative PCR and methylation‑specific PCR were performed to measure the levels of miR‑34b and its promoter methylation in normal cells, eight leukemia cell lines as well as primary leukemic cells isolated from patients newly diagnosed with acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML) and mixed lymphocytic lymphoma. miR‑34b levels in leukemia cell lines and primary leukemic cells were significantly lower than those in normal cells. The miR‑34b promoter was found to be methylated in all leukemia cell lines, 24 of 31 ALL patients and 8 of 19 AML patients, but not in the 23 normal controls. miR‑34b expression and methylation of its promoter were not associated with most clinical parameters assessed; however, miR‑34b levels in prednisone‑sensitive ALL were significantly different from those in insensitive ALL. A cell counting kit‑8 assay showed that transfection of miR‑34b mimics into K562 cells inhibited their proliferation. Furthermore, treatment with the demethylating agent 5‑aza‑2‑deoxycytidine significantly enhanced miR‑34b expression levels and decreased the methylation status of its promoter in HL‑60 and K562 cells. In conclusion, the results of the present study indicated that in pediatric leukemia cells and leukemia cell lines, the expression of miR‑34b is inhibited by methylation of its promoter, which impairs the restraining effects of miR‑34b on cell proliferation. It was also indicated that the expression of miR‑34b in ALL patients may affect their response to early treatments.

摘要

本研究旨在探讨miR-34b启动子甲基化在儿童急性白血病细胞增殖中的作用。采用定量PCR和甲基化特异性PCR检测正常细胞、8种白血病细胞系以及从新诊断的急性淋巴细胞白血病(ALL)、急性髓细胞白血病(AML)和混合淋巴细胞淋巴瘤患者中分离的原发性白血病细胞中miR-34b及其启动子甲基化水平。白血病细胞系和原发性白血病细胞中的miR-34b水平显著低于正常细胞。发现所有白血病细胞系、31例ALL患者中的24例以及19例AML患者中的8例的miR-34b启动子发生甲基化,但23例正常对照中未发生甲基化。miR-34b的表达及其启动子的甲基化与所评估的大多数临床参数无关;然而,泼尼松敏感的ALL患者中的miR-34b水平与不敏感的ALL患者中的miR-34b水平有显著差异。细胞计数试剂盒-8检测显示,将miR-34b模拟物转染到K562细胞中可抑制其增殖。此外,用去甲基化剂5-氮杂-2'-脱氧胞苷处理可显著提高HL-60和K562细胞中miR-34b的表达水平,并降低其启动子的甲基化状态。总之,本研究结果表明,在儿童白血病细胞和白血病细胞系中,miR-34b的表达受到其启动子甲基化的抑制,这削弱了miR-34b对细胞增殖的抑制作用。还表明ALL患者中miR-34b的表达可能影响其对早期治疗的反应。

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