Edalati Fathabad Mahdi, Karimipoor Morteza, Alizadeh Shaban, Abdoli Asghar, Atashi Amir, Sayadi Mahtab
Hematology Department, School of Allied Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Molecular Medicine Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Bioimpacts. 2017;7(2):109-114. doi: 10.15171/bi.2017.14. Epub 2017 Apr 26.
Chronic myelogenous leukemia (CML) is a myeloproliferative disorder caused by the Philadelphia chromosome translocation, at (9; 22), which results in BCR-ABL fusion tyrosine kinase oncoprotein. This fusion induces down-regulation of miR-155. Upregulation of miR-155 can influence cell fate via the effect on p27kip1 and apoptosis. The aim of this study was to induce apoptosis in K562 CML cell line by overexpression of miR-155. The K562 cell line was transfected with pLenti-III-pre mir155-GFP constructs through electroporation. Then, overexpression of miR-155 as well as the expression level of p27kip1 and c-Myc was analyzed by quantitative PCR (qPCR). The level of p27 (Kip1) protein expression was measured by Western blot and the Annexin V method was carried out to investigate apoptosis. Flow cytometric analysis results of K562 cells transfected with pLenti-III-pre mir155-GFP construct showed a significant increase in cell apoptosis. Gene expression and protein level of p27kip1 were upregulated. However, there was no change in c-Myc expression profile. miR-155 could be a promising approach to aid in the treatment of CML. However, further studies are required in this respect.
慢性粒细胞白血病（CML）是一种由9号和22号染色体易位（即费城染色体易位）引起的骨髓增殖性疾病，该易位导致BCR-ABL融合酪氨酸激酶癌蛋白的产生。这种融合导致miR-155的下调。miR-155的上调可通过影响p27kip1和细胞凋亡来影响细胞命运。本研究的目的是通过miR-155的过表达诱导K562慢性粒细胞白血病细胞系凋亡。通过电穿孔法将pLenti-III-pre mir155-GFP构建体转染至K562细胞系。然后，通过定量PCR（qPCR）分析miR-155的过表达以及p27kip1和c-Myc的表达水平。通过蛋白质免疫印迹法检测p27（Kip1）蛋白表达水平，并采用膜联蛋白V法研究细胞凋亡。用pLenti-III-pre mir155-GFP构建体转染K562细胞后的流式细胞术分析结果显示细胞凋亡显著增加。p27kip1的基因表达和蛋白水平上调。然而，c-Myc的表达谱没有变化。miR-155可能是辅助治疗慢性粒细胞白血病的一种有前景的方法。然而，在这方面还需要进一步研究。
