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Evaluation of the Effect of MiR-27a Up-regulation on Megakaryopoiesis in Erythroleukemic K562 Cell Line

Abstract

Author(s): Farshid Karami, Ahmad Kazemi, Fateme Nadali, Shaban Alizade, Ahmad Faramarzi2, Sadegh Alijani1, Neda Minayi, Hamidreza Shahmoradi, Yadi Farshi

"Megakaryopoiesis (platelet production) is a complex process with the potential for regulation at multiple stages. Several studies have indicated that megakaryopoiesis might be regulated post-transcriptionally. This process could be done by small non-coding RNAs called microRNAs which target mRNAs in a sequence-specific manner and lead to translational repression or mRNA decay. The aim of this study is to evaluate the overexpression effects of miR-27a on megakaryocytic differentiation in K562 cell line. K562 cell line was cultured in RPMI 1640 medium, and cloned pre-miR-27a was transfected by lentiviral vector into K562 cells. After RNA extraction and cDNA synthesis in selected days, miRNA up-regulation was confirmed by miRNA real time PCR and then CD41 and CD61 expressions were investigated by RT-PCR. Expression of miR-27a was increased 4.8, 10.3 and 16.8 fold on days 3, 7 and 14 after virus transfect into K562 cell line, in comparison with untransfected cells. Expression level of CD41 and CD61 was positive by RT-PCR technique. The data suggest that miR-27a can be involved in megakaryocytic differentiation and overexpression of megakaryocytic markers in K562 cell line. We suggest that miR-27a may be a significant therapeutic target for increasing both megakaryocytes and platelets in patients with thrombocytopenic diseases."

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