Effects of neurogenin 3 gene silencing on MafA expression in umbilical-derived pancreatic progenitor cells#br#

WU Xing; TAN Meng-Tian; QIN Xiao-Li; HONG Yan

doi:10.16098/j.issn.0529-1356.2020.05.020

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Acta Anatomica Sinica ›› 2020, Vol. 51 ›› Issue (5) : 758-764. DOI: 10.16098/j.issn.0529-1356.2020.05.020

Cell and Molecules Biology

Effects of neurogenin 3 gene silencing on MafA expression in umbilical-derived pancreatic progenitor cells#br#

WU Xing¹ TAN Meng-tian² QIN Xiao-li¹HONG Yan^1*

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Abstract

Objective To investigate the role of neurogenin 3 (Ngn3) gene in the differentiation of human umbilical cord mesenchymal stem cells (hUMSCs) into pancreatic progenitor cells and the effect on Mus musculus v-maf musculoaponeurotic fibrosarcoma oncogene family, protein A(MafA) expression. Methods Isolated, cultured and identified the hUMSCs. Inducing the differentiation of hUMSCs into insulin-secreting cells in stages, and divided them into normal induction group, gene silencing group, and empty virus transfection group.The latter two groups were transfected with interfering virus and empty virus on the 7th day of induction.Detection of infection after puromycin screening was performed. After 21 days of induction,observed the changes of cell morphology by the inverted phase contrast microscope and electron microscope, then detecting the expression of insulin, glucagon and Ngn3 in each group of cells by immunocytochemical technique, detecting the changes of Ngn3 and MafA expression by Real-time PCR and Western blotting. Results The hUMSCs were successfully isolated and cultured.The cells in the normal induction group and the empty virus transfection group were differentiated into pancreatic progenitor cells by stage induction, and Ngn3, Insulin, and glucagon were positively expressed, and secreted particles were observed under electron microscope. Successfully silenced the Ngn3 gene of the gene silencing group of cells, and the cells of this group showed negative expression of Ngn3, insulin and glucagon.Ngn3 and MafA decreased significantly in the gene silencing group (P<0.05). Conclusion Ngn3 gene silencing blocked the differentiation of hUMSCs into insulin-secreting cells and suppressed the expression of MafA.

Key words

Human umbilical cord mesenchymal stem cell / Pancreatic progenitor cell / Neurogenin 3 / Gene silencing / Mus musculus v-maf musculoaponeurotic fibrosarcoma oncogene family, protein A / Real-time PCR

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WU Xing TAN Meng-tian QIN Xiao-li HONG Yan. Effects of neurogenin 3 gene silencing on MafA expression in umbilical-derived pancreatic progenitor cells#br#[J]. Acta Anatomica Sinica. 2020, 51(5): 758-764 https://doi.org/10.16098/j.issn.0529-1356.2020.05.020

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