PDIA3 modulates genomic response to 1,25-dihydroxyvitamin D3 in squamous cell carcinoma of the skin

Joanna I Nowak; Anna M Olszewska; Anna Piotrowska; Kamil Myszczyński; Paweł Domżalski; Michał A Żmijewski

doi:10.1016/j.steroids.2023.109288

PDIA3 modulates genomic response to 1,25-dihydroxyvitamin D₃ in squamous cell carcinoma of the skin

Steroids. 2023 Nov:199:109288. doi: 10.1016/j.steroids.2023.109288. Epub 2023 Aug 5.

Authors

Joanna I Nowak¹, Anna M Olszewska², Anna Piotrowska³, Kamil Myszczyński⁴, Paweł Domżalski⁵, Michał A Żmijewski⁶

Affiliations

¹ Department of Histology, Medical University of Gdansk, 1a Dębinki, 80-211 Gdansk, Poland. Electronic address: j.chorzepa@gumed.edu.pl.
² Department of Histology, Medical University of Gdansk, 1a Dębinki, 80-211 Gdansk, Poland. Electronic address: anna.olszewska@gumed.edu.pl.
³ Department of Histology, Medical University of Gdansk, 1a Dębinki, 80-211 Gdansk, Poland. Electronic address: annapiotrowska@gumed.edu.pl.
⁴ Centre of Biostatistics and Bioinformatics Analysis Medical University of Gdansk, 1a Debinki, 80-211 Gdansk, Poland. Electronic address: kamil.myszczynski@gumed.edu.pl.
⁵ Department of Histology, Medical University of Gdansk, 1a Dębinki, 80-211 Gdansk, Poland. Electronic address: pawel.domzalski@gumed.edu.pl.
⁶ Department of Histology, Medical University of Gdansk, 1a Dębinki, 80-211 Gdansk, Poland. Electronic address: mzmijewski@gumed.edu.pl.

PMID: 37549780
DOI: 10.1016/j.steroids.2023.109288

Abstract

An active form of vitamin D₃ (1,25-dihydroxyvitamin D₃) acts through vitamin D receptor (VDR) initiating genomic response, but several studies described also non-genomic actions of 1,25-dihydroxyvitamin D₃, implying the role of PDIA3 in the process. PDIA3 is a membrane-associated disulfide isomerase involved in disulfide bond formation, protein folding, and remodeling. Here, we used a transcriptome-based approach to identify changes in expression profiles in PDIA3-deficient squamous cell carcinoma line A431 after 1,25-dihydroxyvitamin D₃ treatment. PDIA3 knockout led to changes in the expression of more than 2000 genes and modulated proliferation, cell cycle, and mobility of cells; suggesting an important regulatory role of PDIA3. PDIA3-deficient cells showed increased sensitivity to 1,25-dihydroxyvitamin D₃, which led to decrease migration. 1,25-dihydroxyvitamin D₃ treatment altered also genes expression profile of A431ΔPDIA3 in comparison to A431WT cells, indicating the existence of PDIA3-dependent genes. Interestingly, classic targets of VDR, including CAMP (Cathelicidin Antimicrobial Peptide), TRPV6 (Transient Receptor Potential Cation Channel Subfamily V Member 6), were regulated differently by 1,25-dihydroxyvitamin D₃, in A431ΔPDIA3. Deletion of PDIA3 impaired 1,25-dihydroxyvitamin D₃-response of genes, such as PTGS2, MMP12, and FOCAD, which were identified as PDIA3-dependent. Additionally, response to 1,25-dihydroxyvitamin D₃ in cancerous A431 cells differed from immortalized HaCaT keratinocytes, used as non-cancerous control. Finally, silencing of PDIA3 and 1,25-dihydroxyvitamin D₃, at least partially reverse the expression of cancer-related genes in A431 cells, thus targeting PDIA3 and use of 1,25-dihydroxyvitamin D₃ could be considered in a prevention and therapy of the skin cancer. Taken together, PDIA3 has a strong impact on gene expression and physiology, including genomic response to 1,25-dihydroxyvitamin D₃.

Keywords: 1,25-dihydroxyvitaminvitamin D3; PDIA3; Squamous cell carcinoma; Transcriptome.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carcinoma, Squamous Cell* / genetics
Genomics
Humans
Protein Disulfide-Isomerases* / genetics
Protein Disulfide-Isomerases* / metabolism
Receptors, Calcitriol / genetics
TRPV Cation Channels / metabolism
Vitamin D* / metabolism

Substances

1,25-dihydroxyvitamin D
PDIA3 protein, human
Protein Disulfide-Isomerases
Receptors, Calcitriol
Vitamin D
TRPV6 channel
TRPV Cation Channels