Abstract
Transient receptor potential vanilloid 3 (TRPV3) is highly expressed in skin keratinocytes where it forms Ca2+-permeable nonselective cation channels to regulate various cutaneous functions. TRPV3 expression is upregulated in many skin disorders. Here, we examined how TRPV3 affects keratinocyte proliferation and investigated the underlying mechanism. Topical application of TRPV3 agonist, carvacrol, increased skin thickness in wild type (WT) mice but not in TRPV3 knockout (KO) mice. Carvacrol promoted proliferation of human keratinocytes HaCaT cells at concentrations ≤ 100 μM, but at 300 μM, it decreased cell viability, suggesting a nonmonotonic proliferative effect. Suppression of TRPV3 expression abolished carvacrol-induced cell proliferation while overexpression of TRPV3 enhanced HaCaT cell proliferation. Carvacrol also stimulated Ca2+ influx and proliferation of primary keratinocytes prepared from WT but not TRPV3 KO mice, suggesting that carvacrol-stimulated cell proliferation was dependent on TRPV3-mediated Ca2+ influx. Mechanistic investigation demonstrated that carvacrol stimulated TGFα release and increased phosphorylation levels of EGFR, PI3K, and NF-κB, effects abolished by suppression of TRPV3 expression and CaMKII inhibition. Moreover, inhibition of CaMKII, EGFR, PI3K, or NF-κB diminished carvacrol-induced cell proliferation. We conclude that while strong activation of TRPV3 may cause cell death, moderate activation of TRPV3 promotes cell proliferation in keratinocytes through Ca2+/CaMKII→TGFα/EGFR→PI3K→NF-κB signaling.
Headlights
1. Carvacrol induces epidermal hyperplasia and keratinocyte proliferation.
2. TRPV3 mediates carvacrol-induced epidermal hyperplasia and keratinocyte proliferation.
3. TRPV3 acts through Ca2+/CaMKII→TGFα/EGFR→PI3K→NF-κB signaling to promote keratinocyte proliferation.
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Abbreviations
- 2-APB:
-
2-Aminoethoxydiphenyl borate
- BrdU:
-
5-Bromo-2-deoxyUridine
- BSA:
-
Bovine serum albumin
- CaMKII:
-
Ca2+/calmodulin-dependent protein kinase II
- EGFR:
-
Epidermal growth factor receptor
- EGTA:
-
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid
- FBS:
-
Fetal bovine serum
- GOF:
-
Gain-of-function
- H&E:
-
Hematoxylin and eosin
- Na2EDTA:
-
Ethylenediaminetetraacetic acid disodium salt dihydrate
- NF-κB:
-
Nuclear factor kappa-light-chain-enhancer of activated B cells
- OS:
-
Olmsted Syndrome
- PI3K:
-
Phosphoinositide 3-kinase
- TGFα:
-
Transforming growth factor-α
- TRPV3:
-
Transient receptor potential vanilloid 3
- WT:
-
Wild type
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This work was supported by the National Natural Science Foundation of China (81972960, 21777192); National Science and Technology Major Projects for “Major New Drugs Innovation and Development” (2018ZX09101003-004-002); and the National Key Laboratory of Natural Medicines, China Pharmaceutical University (SKLNMZZCX201825), the “Double First-Class” project by China Pharmaceutical University (CPU2018GY18), and the joint project between China Pharmaceutical University and East South University (2242019K3DZ01).
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Zhengyu Cao and Fang Zhao did the conceptualization. Yujing Wang, Hang Li, and Fang Zhao were responsible for the investigation. Chu Xue, Hao Chen, and Yanning Xue contributed the data analysis. Yujing Wang, Fang Zhao, and Zhengyu Cao contributed in the writing—original draft preparation. Zhengyu Cao and Michael X. Zhu contributed in the writing—review and editing.
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Wang, Y., Li, H., Xue, C. et al. TRPV3 enhances skin keratinocyte proliferation through EGFR-dependent signaling pathways. Cell Biol Toxicol 37, 313–330 (2021). https://doi.org/10.1007/s10565-020-09536-2
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DOI: https://doi.org/10.1007/s10565-020-09536-2