PMID- 31749426 OWN - NLM STAT- MEDLINE DCOM- 20210609 LR - 20210609 IS - 1875-5666 (Electronic) IS - 1566-5240 (Linking) VI - 20 IP - 4 DP - 2020 TI - Overexpression of Pygo2 Increases Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Cardiomyocyte-like Cells. PG - 318-324 LID - 10.2174/1566524019666191017150416 [doi] AB - BACKGROUND: Our previous studies have shown that Pygo (Pygopus) in Drosophila plays a critical role in adult heart function that is likely conserved in mammals. However, its role in the differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) into cardiomyocytes remains unknown. OBJECTIVE: To investigate the role of pygo2 in the differentiation of hUC-MSCs into cardiomyocytes. METHODS: Third passage hUC-MSCs were divided into two groups: a p+ group infected with the GV492-pygo2 virus and a p- group infected with the GV492 virus. After infection and 3 or 21 days of incubation, Quantitative real-time PCR (qRT-PCR) was performed to detect pluripotency markers, including OCT-4 and SOX2. Nkx2.5, Gata-4 and cTnT were detected by immunofluorescence at 7, 14 and 21 days post-infection, respectively. Expression of cardiac-related genes-including Nkx2.5, Gata-4, TNNT2, MEF2c, ISL-1, FOXH1, KDR, alphaMHC and alpha-Actin-were analyzed by qRT-PCR following transfection with the virus at one, two and three weeks. RESULTS: After three days of incubation, there were no significant changes in the expression of the pluripotency stem cell markers OCT-4 and SOX2 in the p+ group hUC-MSCs relative to controls (OCT-4: 1.03 +/- 0.096 VS 1, P > 0.05, SOX2: 1.071 +/- 0.189 VS 1, P > 0.05); however, after 21 days, significant decreases were observed (OCT-4: 0.164 +/- 0.098 VS 1, P < 0.01, SOX2: 0.209 +/- 0.109 VS 1, P < 0.001). Seven days following incubation, expression of mesoderm specialisation markers, such as Nkx2.5, Gata-4, MEF2c and KDR, were increased; at 14 days following incubation, expression of cardiac genes, such as Nkx2.5, Gata-4, TNNT2, MEF2c, ISL-1, FOXH1, KDR, alphaMHC and alpha-Actin, were significantly upregulated in the p+ group relative to the p- group (P < 0.05). Taken together, these findings suggest that overexpression of pygo2 results in more hUCMSCs gradually differentiating into cardiomyocyte-like cells. CONCLUSION: We are the first to show that overexpression of pygo2 significantly enhances the expression of cardiac-genic genes, including Nkx2.5 and Gata-4, and promotes the differentiation of hUC-MSCs into cardiomyocyte-like cells. CI - Copyright(c) Bentham Science Publishers; For any queries, please email at epub@benthamscience.net. FAU - Yang, Lei AU - Yang L AD - Department of Cardiac Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. FAU - Zhu, Shuoji AU - Zhu S AD - Department of Cardiac Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. FAU - Li, Yongqing AU - Li Y AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China. FAU - Zhuang, Jian AU - Zhuang J AD - Department of Cardiac Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. FAU - Chen, Jimei AU - Chen J AD - Department of Cardiac Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. FAU - Huang, Huanlei AU - Huang H AD - Department of Cardiac Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. FAU - Chen, Yu AU - Chen Y AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China. FAU - Wen, Yulin AU - Wen Y AD - Department of Cardiac Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. FAU - Wen, Yao AU - Wen Y AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China. FAU - Guo, Huiming AU - Guo H AD - Department of Cardiac Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. FAU - Fan, Xiongwei AU - Fan X AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China. FAU - Yuan, Wuzhou AU - Yuan W AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China. FAU - Jiang, Zhigang AU - Jiang Z AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China. FAU - Wang, Yuequn AU - Wang Y AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China. FAU - Wu, Xiushan AU - Wu X AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China. FAU - Zhu, Ping AU - Zhu P AD - Department of Cardiac Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - Netherlands TA - Curr Mol Med JT - Current molecular medicine JID - 101093076 RN - 0 (Biomarkers) RN - 0 (GATA4 Transcription Factor) RN - 0 (Homeobox Protein Nkx-2.5) RN - 0 (Intracellular Signaling Peptides and Proteins) RN - 0 (NKX2-5 protein, human) RN - 0 (PYGO2 protein, human) RN - 0 (Troponin T) SB - IM MH - Biomarkers/metabolism MH - *Cell Differentiation MH - Cell Proliferation MH - Cell Separation MH - GATA4 Transcription Factor/metabolism MH - Gene Expression Regulation MH - Homeobox Protein Nkx-2.5/metabolism MH - Humans MH - Intracellular Signaling Peptides and Proteins/*metabolism MH - Mesenchymal Stem Cells/*cytology MH - Myocytes, Cardiac/*cytology MH - Organogenesis/genetics MH - Pluripotent Stem Cells/metabolism MH - Transcription, Genetic MH - Troponin T/metabolism MH - Umbilical Cord/*cytology MH - Up-Regulation OTO - NOTNLM OT - Human umbilical cord mesenchymal stem cells OT - Nkx2.5 OT - SOX2 OT - cardiomyocytes OT - differentiation OT - overexpression OT - pygo2. EDAT- 2019/11/22 06:00 MHDA- 2021/06/10 06:00 CRDT- 2019/11/22 06:00 PHST- 2019/07/17 00:00 [received] PHST- 2019/09/25 00:00 [revised] PHST- 2019/09/26 00:00 [accepted] PHST- 2019/11/22 06:00 [pubmed] PHST- 2021/06/10 06:00 [medline] PHST- 2019/11/22 06:00 [entrez] AID - CMM-EPUB-101581 [pii] AID - 10.2174/1566524019666191017150416 [doi] PST - ppublish SO - Curr Mol Med. 2020;20(4):318-324. doi: 10.2174/1566524019666191017150416.