The key regulation of LncRNA MALAT1/miR-124-3p/PI3K axis during reprogramming of primary mouse hepatocytes into insulin-producing cells

In this study, a considerable augmentation in the expression of the endogenous long noncoding RNA MALAT1 (lncRNA MALAT1) and PI3K was discerned, in contrast to a significant diminution in miR-124-3p expression, during the progression of in vitro induced differentiation of primary mouse hepatocytes. Bioinformatics scrutiny and the luciferase reporter assay unveiled the binding of miR-124-3p to specific loci in lncRNA MALAT1 and PI3K, suppressing their expression. Elevated expression of lncRNA MALAT1 potently diminished the expression levels of PI3K and attenuated the efficacy of primary mouse hepatocytes differentiation into insulin-producing cells. Furthermore, the in vitro responsiveness of primary mouse hepatocytes-derived β islet-like cells to high glucose stimuli and insulin release by these cells were significantly diminished in the overexpression of lncRNA MALAT1 transfection group, while the outcomes were converse in the si-lncRNA MALAT1 group. The experimental findings suggest that lncRNA MALAT1 effectively sustains PI3K gene expression through competitive binding to miR-124-3p, accomplishing regulation of primary mouse hepatocytes directed β islet-like cell differentiation efficiency. To differentiate the primary mouse hepatocytes into IPCs using our step-wise protocol,We further extracted the RNA from various stages cells, and performed a high throughput RNA-seq analysis, upon dissecting differentially expressed mRNA and lncRNA,