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LncRNA-uc.40 silence promotes P19 embryonic cells differentiation to cardiomyocyte via the PBX1 gene

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Abstract

Uc.40 is a long noncoding RNA that is highly conserved among different species, although its function is unknown. It is highly expressed in abnormal human embryonic heart. We previously reported that overexpression of uc.40 promoted apoptosis and inhibited proliferation of P19 cells, and downregulated PBX1, which was identified as a potential target gene of uc.40. The current study evaluated the effects of uc40-siRNA-44 (siRNA against uc.40) on the differentiation, proliferation, apoptosis, and mitochondrial function in P19 cells, and investigated the relationship between uc.40 and PBX1 in cardiomyocytes. The uc.40 silencing expression was confirmed by quantitative real-time polymerase chain reaction (RT-PCR). Observation of morphological changes in transfected P19 cells during different stages of differentiation revealed that uc40-siRNA-44 increased the number of cardiomyocyes. There was no significant difference in the morphology or time of differentiation between the uc40-siRNA-44 group and the control group. uc40-siRNA-44 significantly promoted proliferation of P19 cells and inhibited serum starvation-induced apoptosis. There was no significant difference in mitochondrial DNA copy number or cellular ATP level between the two groups, and ROS levels were significantly decreased in uc40-siRNA-44-transfected cells. The levels of PBX1 and myocardial markers of differentiation were examined in transfected P19 cells; uc40-siRNA-44 downregulated myocardial markers and upregulated PBX1 expression. These results suggest that uc.40 may play an important role during the differentiation of P19 cells by regulation of PBX1 to promote proliferation and inhibit apoptosis. These studies provide a foundation for further study of uc.40/PBX1 in cardiac development.

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Acknowledgments

This study was supported by grants from the National Natural Science Foundation of China (grant no. 81470376), the National Natural Science Foundation of Jiangsu Province of China (grant no. BK20141077), Changzhou Municipal Science and Technology Bureau guidance project (grant no. 2016361), and Changzhou Science and Technique Development Foundation (grant no. 20170397).

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Authors and Affiliations

  1. Medical Research Center, The Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University, Changzhou, 213003, China

    Rongqiang Wu, Yu Wan & Shizhong Wang

  2. Department of Pediatrics, Changzhou Children’s Hospital, Nantong Medical University, Nantong City, China

    Peng Xue & Meng Gu

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  1. Rongqiang Wu
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  2. Peng Xue
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  3. Yu Wan
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  4. Shizhong Wang
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  5. Meng Gu
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Correspondence to Shizhong Wang or Meng Gu.

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Editor: Tetsuji Okamoto

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Wu, R., Xue, P., Wan, Y. et al. LncRNA-uc.40 silence promotes P19 embryonic cells differentiation to cardiomyocyte via the PBX1 gene. In Vitro Cell.Dev.Biol.-Animal 54, 600–609 (2018). https://doi.org/10.1007/s11626-018-0284-0

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  • DOI: https://doi.org/10.1007/s11626-018-0284-0

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