Expression of Cell Pyroptosis-Associated TLR4 Signalling Pathway During the Progression from Actinic Keratosis to Cutaneous Squamous Cell Carcinoma

Xiaoyu ZHAO; Minyan ZHANG; Huiya CHEN; Tingting CUI; Yuhan HUANG; Dan XU

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Xiaoyu ZHAO, Minyan ZHANG, Huiya CHEN, Tingting CUI, Yuhan HUANG, Dan XU. Expression of Cell Pyroptosis-Associated TLR4 Signalling Pathway During the Progression from Actinic Keratosis to Cutaneous Squamous Cell Carcinoma[J]. Journal of Kunming Medical University.

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Expression of Cell Pyroptosis-Associated TLR4 Signalling Pathway During the Progression from Actinic Keratosis to Cutaneous Squamous Cell Carcinoma

1.
Department of Dermatology，The First Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650032
2.
Department of Dermatology，Dongguan Liaobu Hospital，Dongguan Guangdong 523400，China

Received Date: 2025-04-22

Abstract

Abstract

Objective To investigate whether the pyroptosis-related Toll-like receptor 4 （TLR4） signaling pathway influences the malignant transformation of actinic keratosis （AK） into cutaneous squamous cell carcinoma （SCC）. Methods A total of 6 lesion tissue samples each from patients with AK and SCC, as well as 5 normal skin tissue samples from healthy subjects, were collected from the First Affiliated Hospital of Kunming Medical University between August 2020 and August 2021 as controls. Quantitative PCR （qPCR） and Western blot analysis were performed to determine the mRNA and protein expression levels of TLR4 pathway–related factors, including TLR4, CPB1, NLRP3, IL-1β, and IL-18. TLR4/TUNEL double immunofluorescence staining was used to evaluate TLR4 expression and the level of cellular pyroptosis in tissue samples. In addition, Western blotting was performed to analyze the expression differences of pyroptosis-related core proteins （pro-caspase-1, cleaved caspase-1/p20, GSDMD, and cleaved N-terminal GSDMD） and TLR4 among the normal keratinocyte cell line HaCaT and SCC cell lines A431 and SCL-1. Results Quantitative PCR and Western blot results showed that the mRNA and protein expression levels of TLR4, CPB1, NLRP3, IL-1β, and IL-18 were significantly higher in SCC tissues than in AK and normal skin tissues （P < 0.05）. TLR4/TUNEL double immunofluorescence results revealed a progressive increasing trend in TLR4 expression and pyroptosis levels from normal skin to AK and further to SCC （P < 0.05）. Furthermore, in SCL-1 cells, the expression levels of pro-caspase-1, cleaved caspase-1/p20, cleaved N-terminal GSDMD, and TLR4 were significantly upregulated （P < 0.05）, whereas in A431 cells, only TLR4 expression was increased, and the levels of other pyroptosis-related proteins were downregulated compared to HaCaT cells （P < 0.05）. Conclusion The TLR4 signaling pathway is highly expressed in both AK and SCC, with higher levels observed in SCC, suggesting that it may promote the malignant transformation of AK into SCC. Moreover, the activation status of the TLR4-mediated pyroptosis pathway differs among SCC cell lines: SCL-1 cells exhibit robust pyroptotic activation, whereas A431 cells display upregulated TLR4 expression but suppressed pyroptosis. These findings indicate that TLR4-mediated pyroptosis may play a crucial regulatory role in the pathogenesis and progression of cutaneous squamous cell carcinoma and in the malignant transformation of its precancerous lesions.
- Cutaneous squamous cell carcinoma,
- Actinic keratosis,
- Cell pyroptosis,
- TLR4

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References(33)

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