2022, Vol. 26 ›› Issue (30): 4847-4851

Role of long non-coding RNA nuclear-enriched abundant transcript 1 in the differentiation of human umbilical cord mesenchymal stem cells into hepatocytes

Wang Dandan1, Yu Yabin2, Liu Shiqi1, Yan Yulou1, Zhang Jianhuai2   

Abstract: BACKGROUND: Human umbilical cord mesenchymal stem cells can be induced to differentiate into hepatocytes with the help of cytokines in vitro, but the resulting hepatoid cells are not yet functionally mature. It has been gradually found that long non-coding RNA (lncRNA) is closely related to cell differentiation and the purpose of promoting cell differentiation is achieved by interfering with the expression of some lncRNAs.  
OBJECTIVE: To explore the role of long non-coding RNA nuclear-enriched abundant transcript 1 (lncRNA NEAT1) in the differentiation of human umbilical cord mesenchymal stem cells into hepatocytes.
METHODS:   A differentiational cell line system of human umbilical cord mesenchymal stem cells into hepatocytes was established. Periodic Acid-Schiff staining and indocyanine green intake test were used to assess hepatocellular functional characteristics of the cells. Quantitative real-time reverse transcription polymerase chain reaction was used to detect the expression of hepatocyte specific genes and lncRNA NEAT1. The lentivirus was constructed to interfere with NEAT1 expression, and stably transfected cell lines were obtained 7 days after transfection, and hepatotropic differentiation was induced for 4 weeks. A negative control group for transfection with empty plasmids and a control group without virus transfection were set up. The expression levels of hepatocyte specific genes and proteins were detected by quantitative real-time reverse transcription polymerase chain reaction and western blot assay.  
RESULTS AND CONCLUSION: (1) After Periodic Acid-Schiff staining, there were red and purple glycogen particles in the cytoplasm of cells, and a large number of indocyanine green positive cells were observed after indocyanine green ingestion. (2) With the extension of induction time, the mRNA expression levels of albumin and cytochrome enzyme P450 3A4 mRNA increased gradually, while the mRNA expression levels of NEAT1 decreased gradually. (3) Compared with the negative control group and the control group, the mRNA and protein expression levels of albumin and cytochrome enzyme P450 3A4 were significantly increased after the interference of NEAT1. (4) The results confirm that lncRNA NEAT1 may be involved in the differentiation of human umbilical cord mesenchymal stem cells into hepatocytes.
Key words: long non-coding RNA NEAT1, mesenchymal stem cell, hepatocyte, differentiation, hepatocyte specific genes, transfection