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Characterization of In Vitro Differentiation of Human Primary Keratinocytes by RNA-Seq Analysis
Published on: May 16, 2020
Identification of novel keratinocyte differentiation modulating compounds by high-throughput screening
Masaru Honma1, Mark Stubbs, Ian Collins
1Keratinocyte Laboratory, Cancer Research UK London Research Institute, London.
Journal of Biomolecular Screening
|November 10, 2006
Summary
Researchers screened compounds to find those affecting keratinocyte differentiation. One novel compound promoted differentiation and inhibited cancer cell growth, showing potential for antitumor therapy.
Area of Science:
- Cell Biology
- Dermatology
- Pharmacology
Background:
- Primary human epidermal keratinocytes undergo terminal differentiation.
- Identifying compounds that modulate keratinocyte differentiation is crucial for understanding skin biology and developing therapies.
- High-throughput screening offers a method to discover such compounds efficiently.
Purpose of the Study:
- To identify small molecules that promote or inhibit terminal differentiation of primary human epidermal keratinocytes.
- To discover novel compounds with potential therapeutic applications in skin conditions and cancer.
Main Methods:
- Utilized high-throughput screening of approximately 4000 diverse compounds.
- Employed an In-Cell Western system with immunofluorescent staining for the terminal differentiation marker, involucrin.
- Investigated the effects of known pathway inhibitors (e.g., protein kinase C, protein kinase A, MEK, histone deacetylase inhibitors) and novel compounds.
Main Results:
- Staurosporine and H89 promoted involucrin expression, indicating differentiation.
- U0126, SAHA, and SBHA reduced involucrin expression during calcium-induced stratification.
- Identified one novel compound that induced keratinocyte differentiation and two that inhibited it.
- The differentiation-inducing compound inhibited squamous cell carcinoma growth by promoting differentiation and apoptosis at lower concentrations than in primary keratinocytes.
Conclusions:
- Specific kinase and histone deacetylase inhibitors differentially regulate keratinocyte terminal differentiation.
- A novel small molecule was identified that induces keratinocyte differentiation and exhibits potent antitumor activity against squamous cell carcinoma.
- This differentiation-inducing compound holds promise for developing new anticancer therapies.
