Reports of Biochemistry and Molecular Biology

en Effect of miR-18a-5p, miR-19a-3p, and miR-20a-5p on In Vitro Cardiomyocyte Differentiation of Human Endometrium Tissue-Derived Stem Cells Through Regulation of Smad4 Expression زیست شناسی سلولی Cell Biology مقالات اصلی Original Article <div style="text-align: justify;">Background: Smad4 regulates the expression of the genes required for heart homeostasis. Regarding the central role of microRNAs in cardiac biology, we investigated the expression of the three Smad4-targeting miRNAs, namely miR-18a-5p, miR-19a-3p, and miR-20a-5p, as well as Smad4 during differentiation of human endometrium-derived mesenchymal stem cells (hEMSCs) into cardiomyocytes (CMs). Methods: To evaluate mesenchymal phenotype and multi-lineage differentiation ability of hEMSCs, immunophenotyping by flow cytometry and differentiation into osteoblasts and adipocytes were performed, respectively. For transdifferentiation into CMs, hEMSCs were exposed to a cardiomyogenic medium composed of 5-aza and bFGF for 30 days. The comparison between transcriptional expression levels of Nkx2-5, GATA4, Smad4, TNNT2, TBX5, miR-18a-5p, miR-19a-3p, and miR-20a-5p by qRT-PCR, as well as protein levels of Nkx2-5, Smad4, and cTnT by immunofluorescence staining, was conducted in every 6 days. Results: In vitro, the mesenchymal stem cell phenotype of hEMSCs and their potency for differentiation into other MSCs were confirmed. Differentiated hEMSCs had morphological characteristics of CMs. The percentage of positive cells for Nkx2-5, Smad4, and cTnT proteins was increased following induction and culminated on the 24th day. Also, mRNA levels of Nkx2-5, GATA4, Smad4, TNNT2, and TBX5 exhibited the same trend. The expression of investigated miRNAs was significantly decreased sequentially. A significant negative correlation between expressions of Smad4 and investigated miRNAs was observed. Conclusions: Our results indicate that miR-18a-5p, miR-19a-3p, and miR-20a-5p are involved in the cardiac differentiation propensity of hEMSCs potentially by regulation of Smad levels. Although, more mechanistic experiments are required to confirm this idea.</div> Endometrium-derived mesenchymal stem cells (EMSCs), miR-18a-5p, miR-19a-3p, miR-20a-5p, Smad4. 136 146 http://rbmb.net/browse.php?a_code=A-10-1217-1&slc_lang=en&sid=1 Behnaz Maleki 100319475328460017859 100319475328460017859 No Physiology Research Center, Kashan University of Medical Sciences, Kashan, Iran. Mahdi Noureddini 100319475328460017860 100319475328460017860 No Physiology Research Center, Kashan University of Medical Sciences, Kashan, Iran & Department of Applied Cell Sciences, Faculty of Medicine, Kashan University of Medical Sciences, Kashan, Iran. Somayeh Saadat 100319475328460017861 100319475328460017861 No Physiology Research Center, Kashan University of Medical Sciences, Kashan, Iran. Javad Verdi 100319475328460017862 100319475328460017862 No Department of Applied Cellular Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran. Alireza Farrokhian 100319475328460017863 100319475328460017863 No Department of Cardiology, School of Medicine, Shahid Beheshti Hospital, Kashan University of Medical Sciences, Kashan, Iran. Hossein Ghanbarian 100319475328460017864 100319475328460017864 No Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Ebrahim Cheraghi 100319475328460017865 100319475328460017865 No Department of Biology, Faculty of Science, University of Qom, Qom, Iran. Behrang Alani alani-be@kaums.ac.ir 100319475328460017866 100319475328460017866 Yes Department of Applied Cell Sciences, Faculty of Medicine, Kashan University of Medical Sciences, Kashan, Iran.