Abstract
Adipose-derived stem cells (ADSCs) are an abundant cell source and provide an easy way to harvest mesenchymal stem cells, which are the focus of considerable attention in regenerative medicine. Electric fields (EF) play roles in many biological events and have been reported to promote cell proliferation, migration, and differentiation. In this study, ADSCs were treated with a direct current electric field (DCEF) of either 0 (control group) or 300 mV/mm (EF group) for six hours. RNA screening and analysis revealed that 66, 164, 26, and 1310 circRNAs, lncRNAs, miRNAs, and mRNAs, respectively, were differentially expressed in the DCEF-treated ADSCs compared with untreated ADSCs. Differentially expressed mRNAs were enriched in the MAPK signaling pathway, TNF signaling pathway, and some other pathways. ANXA1, ERRFI1, JAG1, EPHA2, PRR9, and H2AFY2 were related to the keratinocyte differentiation process. Competing endogenous RNA (ceRNA) networks were constructed on the basis of genes in the MAPK signaling pathway. Twenty-one RNAs in the above networks were randomly chosen, and their expression was validated using qRT-PCR, which showed the same expression trends as the RNA sequencing analysis. The MAPK signaling pathway is of great importance in the ADSC processes that occur in a DCEF, including keratinocyte differentiation. Several ceRNAs may participate in the regulation of MAPK signaling. This study may give new insight into the proliferation, migration, and differentiation of ADSCs, which will be valuable for tissue engineering and regenerative medicine.
Highlights
Mesenchymal stem cells (MSCs) are a population of adult stem cells with self-renewal properties and multilineage differentiation capacity
Previous studies have shown that direct current electric field (DCEF) can trigger morphological changes in Adipose-derived stem cells (ADSCs) and force cells to align vertically to the Electric fields (EF) vector or even induce them to migrate to the cathode, which is referred to as galvanotaxis [8]
Differential Expression Analysis of circRNAs, long noncoding RNAs (lncRNAs), miRNAs, and mRNAs. ncRNAs and mRNAs that were differentially expressed in cells of the DCEF group (n = 3) relative to those in the control group are indicated in a heat map and a volcano map (Figure 2)
Summary
Mesenchymal stem cells (MSCs) are a population of adult stem cells with self-renewal properties and multilineage differentiation capacity. Adipose-derived stem cells (ADSCs) are advantageous due to their abundant cell sources and ease of harvesting [4]. Previous studies have shown that DCEF can trigger morphological changes in ADSCs and force cells to align vertically to the EF vector or even induce them to migrate to the cathode, which is referred to as galvanotaxis [8]. There have been no studies on transcriptome sequencing and ceRNA network prediction of ADSCs in an EF. We used whole-transcriptome sequencing technology to investigate the RNA expression profile in ADSCs stimulated by DCEF of 0 and 300 mV/mm for 6 h, respectively. Expressed mRNAs, miRNAs, lncRNAs, and circRNAs were identified. The differentially expressed mRNAs were primarily enriched in the MAPK and TNF signaling pathways. 21 RNAs were randomly chosen, and their expression profiles were validated using qRT-PCR
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