Related Experiment Video
Updated: Jun 1, 2025
Clonogenic Assay: Adherent Cells
Published on: March 13, 2011
Fluoxetine exerts anti-proliferative effect in human epidermal keratinocytes
Dorottya Ádám1,2, József Arany1,2, Kinga Fanni Tóth1,2
1Department of Physiology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
Abstract:
We have recently shown that fluoxetine (FX) suppressed polyinosinic-polycytidylic acid-induced inflammatory response and endothelin release in human epidermal keratinocytes, via the indirect inhibition of the phosphoinositide 3-kinase (PI3K)-pathway. Because PI3K-signaling is a positive regulator of the proliferation, in the current, highly focused follow-up study, we assessed the effects of FX (14 µM) on the proliferation and differentiation of human epidermal keratinocytes. We found that FX exerted anti-proliferative actions in 2D cultures (HaCaT and primary human epidermal keratinocytes [NHEKs]; 48- and 72-h; CyQUANT-assay) as well as in 3D reconstructed epidermal equivalents (48-h; Ki-67 immunohistochemistry). Importantly, FX did not influence epidermal thickness (hematoxylin-eosin staining), and it did not have a major impact on the differentiation-associated alteration of the gene expression pattern (24-h treatments; RNA-Seq). Moreover, neither keratin (K)-1, nor K10 expression was altered by FX in NHEKs (RT-qPCR) or in 3D epidermal equivalents (semi-quantitative immunohistomorphometry). FX did not influence differentiation-induced up-regulation of occludin (RT-qPCR; NHEKs), and did not alter differentiation-associated barrier forming capacity of epidermal keratinocytes (electrical impedance; Lucifer Yellow penetration assay). Our data indicate that, besides the previously reported combined anti-inflammatory and putative anti-pruritic effects, FX may also suppress proliferation of human epidermal keratinocytes without impairing their differentiation and barrier-forming capacity.
Insights
Fluoxetine (FX) suppresses human epidermal keratinocyte proliferation without affecting differentiation or barrier function. This study investigated FX
Area of Science:
- Dermatology
- Cell Biology
- Pharmacology
Background:
- Fluoxetine (FX) previously demonstrated anti-inflammatory effects in human epidermal keratinocytes.
- Phosphoinositide 3-kinase (PI3K) signaling regulates keratinocyte proliferation.
- The impact of FX on keratinocyte proliferation and differentiation remained unexamined.
Purpose of the Study:
- To investigate the effects of fluoxetine (FX) on the proliferation and differentiation of human epidermal keratinocytes.
- To determine if FX impairs epidermal differentiation or barrier formation.
Main Methods:
- Assessed FX effects on keratinocyte proliferation using CyQUANT assays in 2D cultures (HaCaT, NHEKs) and Ki-67 immunohistochemistry in 3D epidermal equivalents.
- Analyzed epidermal thickness via hematoxylin-eosin staining.
- Evaluated gene expression patterns related to differentiation using RNA-Seq and RT-qPCR (K1, K10, occludin).
- Assessed barrier function using electrical impedance and Lucifer Yellow penetration assays.
Main Results:
- FX (14 µM) exhibited anti-proliferative effects in both 2D and 3D keratinocyte cultures.
- FX did not significantly alter epidermal thickness or differentiation-associated gene expression.
- Keratin 1, keratin 10, and occludin expression remained unchanged by FX treatment.
- FX did not impair the differentiation-associated barrier-forming capacity of epidermal keratinocytes.
Conclusions:
- Fluoxetine suppresses human epidermal keratinocyte proliferation.
- FX does not negatively impact keratinocyte differentiation or barrier function.
- FX presents potential therapeutic applications for skin conditions involving inflammation and hyperproliferation.
