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Updated: Feb 23, 2026
Characterization of In Vitro Differentiation of Human Primary Keratinocytes by RNA-Seq Analysis
Published on: May 16, 2020
Differential miRNA expression profiles in human keratinocytes in response to protein kinase C inhibitor
Yang Liu1, Liangchen Zhong2, Dewu Liu1
1Burns Center, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Abstract:
Aberrant expression of microRNAs (miRNAs) is widely accepted to be involved in keratinocyte differentiation and to be dependent on activation of the protein kinase C (PKC) pathway. However, the miRNA profiles and biological characteristics of keratinocytes induced by specific inhibitors of PKC have yet to be elucidated. The present study aimed to explore the differential miRNA expression profiles in keratinocytes treated with the PKC inhibitor GF109203X, by conducting a bioinformatics analysis. Parts of the GF109203X‑induced keratinocytes formed distinct clones after 2 days of culture, and the expression of intergrin β1, cytokeratin (CK)19 and CK14 were positive, whereas CK10 expression was negative. A total of 79 miRNAs were differentially expressed in keratinocytes treated with GF109203X, among which 45 miRNAs were upregulated and 34 were downregulated. The significantly upregulated microRNAs includedhsa‑miR‑1‑3p and miR‑181c‑5p, whereas hsa‑miR‑31‑5p and hsa‑let‑7c‑3p were significantly downregulated. In addition, the results of reverse transcription‑quantitative polymerase chain reaction exhibited consistency with the microarray results. An enrichment analysis demonstrated that certain target genes of the differentially expressed miRNAs serve an important role in cell proliferation and differentiation, cell cycle progression and apoptosis, etc. These results revealed that GF109203X induced the differential expression of certain miRNAs when keratinocytes began showing the characteristics of epidermal‑like stem cells, which may provide a novel approach for wound healing and regeneration of skin tissues.
Insights
Protein kinase C (PKC) inhibition alters microRNA (miRNA) profiles in keratinocytes, inducing epidermal-like stem cell characteristics. This discovery offers new avenues for skin tissue regeneration and wound healing strategies.
Area of Science:
- Dermatology
- Molecular Biology
- Biotechnology
Background:
- Aberrant microRNA (miRNA) expression is linked to keratinocyte differentiation.
- The protein kinase C (PKC) pathway influences miRNA expression.
- The specific miRNA profiles induced by PKC inhibitors in keratinocytes remain largely uncharacterized.
Purpose of the Study:
- To investigate differential miRNA expression profiles in keratinocytes treated with the PKC inhibitor GF109203X.
- To analyze the biological characteristics of these GF109203X-induced keratinocytes.
- To explore the potential of these findings for skin regeneration.
Main Methods:
- Bioinformatics analysis of miRNA expression in GF109203X-treated keratinocytes.
- Cell culture and observation of keratinocyte clonal formation.
- Assessment of specific protein markers (integrin β1, CK19, CK14, CK10) via expression analysis.
- Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) for validation.
- Enrichment analysis of miRNA target genes.
Main Results:
- GF109203X treatment induced keratinocytes with positive expression of integrin β1, CK19, and CK14, and negative for CK10, resembling epidermal-like stem cells.
- A total of 79 miRNAs were differentially expressed: 45 upregulated and 34 downregulated.
- Key upregulated miRNAs included hsa-miR-1-3p and miR-181c-5p; downregulated miRNAs included hsa-miR-31-5p and hsa-let-7c-3p.
- RT-qPCR results corroborated microarray findings.
- Enrichment analysis indicated that target genes of differentially expressed miRNAs are involved in cell proliferation, differentiation, cell cycle, and apoptosis.
Conclusions:
- GF109203X induces differential miRNA expression in keratinocytes, promoting epidermal-like stem cell characteristics.
- These miRNA changes are associated with critical cellular processes like proliferation and differentiation.
- The study suggests a novel approach for wound healing and skin tissue regeneration by modulating PKC and miRNA pathways.
