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Inducing Apoptosis and Decreasing Cell Proliferation in Human Acute Promyelocytic Leukemia Through Regulation Expression of CASP3 by Let-7a-5p Blockage

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Indian Journal of Hematology and Blood Transfusion Aims and scope Submit manuscript

Abstract

MicroRNAs (miRNAs) are short and single strand non-coding RNAs that involved in post-transcriptional regulation of gene expression. Dysregulation of miRNA expression is important event in the many of malignant diseases. Up-regulation of Let-7a-5p expression in acute myeloid leukemia in human in previous studies was reported. In this study blockage of Let-7a-5p in human acute promyelocytic leukemia cell line (HL60) was done by using locked nucleic acid (LNA) method and subsequently expression of Let-7a-5p, cell proliferation, apoptosis, necrosis, and CASP3 expression was measured. At three time points 24, 48 and 72 h after LNA anti- Let-7a-5p transfection, assessment of Let-7a-5p expression by qRT real-time PCR was completed. The MTT assay and annexin/PI staining have been performed. Also, CASP3 expression at different time points after LNA anti-Let-7a-5p transfection in HL60 cell line was measured. The results at three-time points after LNA transfection were represented that Let-7a-5p expression was lower in the LNA-anti-Let-7a group compared to the control groups. The cell viability significantly was different between LNA-anti-Let-7a group and control groups. Increasing apoptotic ratio was associated with Let-7a-5p blockage in the LNA-anti-Let-7a group compared with control groups. Also, the necrotic ratio was higher in the LNA-anti-Let-7a group rather than the other groups. Western blotting revealed that CASP3 expression associated with Let-7a-5p inhibition. Our results displayed that blockage of Let-7a-5p can reduced cell viability mainly due to the induction of apoptosis and CASP3 up-regulation in HL60 cells. These results can be useful in translational medicine for research of antisense therapy in leukemia.

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Acknowledgements

This study was conducted with the support of Isfahan University of Medical Sciences (IRAN).

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Authors and Affiliations

  1. Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

    Mahdi Fasihi-Ramandi, Abbas Moridnia & Ali Najafi

  2. Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Science, Isfahan, 81744-176, Iran

    Mohammadreza Sharifi

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  1. Mahdi Fasihi-Ramandi
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  2. Abbas Moridnia
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  3. Ali Najafi
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  4. Mohammadreza Sharifi
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Correspondence to Mohammadreza Sharifi.

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This study was approved by the local ethics committee of Isfahan University of Medical Sciences (IRAN) and the studies have been approved by the appropriate institutional and/or a national research ethics committee and have been performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

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Fasihi-Ramandi, M., Moridnia, A., Najafi, A. et al. Inducing Apoptosis and Decreasing Cell Proliferation in Human Acute Promyelocytic Leukemia Through Regulation Expression of CASP3 by Let-7a-5p Blockage. Indian J Hematol Blood Transfus 34, 70–77 (2018). https://doi.org/10.1007/s12288-017-0809-9

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  • DOI: https://doi.org/10.1007/s12288-017-0809-9

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