LncRNA-ZNF252P-AS1/miR-15b-5p promotes the proliferation of keloid fibroblast by regulating the BTF3-STAT3 signaling pathway

Yu Guo; Mengjuan Li; Jianhong Long; Pengju Fan; Chenchen Zuo; Yongjie Wang

doi:10.1016/j.jdermsci.2022.12.010

LncRNA-ZNF252P-AS1/miR-15b-5p promotes the proliferation of keloid fibroblast by regulating the BTF3-STAT3 signaling pathway

J Dermatol Sci. 2022 Dec;108(3):146-156. doi: 10.1016/j.jdermsci.2022.12.010. Epub 2022 Dec 29.

Authors

Yu Guo¹, Mengjuan Li², Jianhong Long², Pengju Fan², Chenchen Zuo², Yongjie Wang³

Affiliations

¹ Department of Burns and Plastic Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China.
² Department of Burns and Plastic Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China.
³ Department of Burns and Plastic Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China. Electronic address: [email protected].

PMID: 36641250
DOI: 10.1016/j.jdermsci.2022.12.010

Abstract

Background: JAK2/STAT3 signaling pathway plays an important role in keloid formation, but the upstream mechanism of their activation remains unclear.

Objective: This study aims to investigate the possible mechanism of lncRNA-ZNF252P-AS1 in keloid.

Methods: The differentially expressed genes in keloid and their upstream regulatory miRNAs and long non-coding RNAs (lncRNAs) were analyzed by bioinformatics database, and the targeting relationship was further verified by dual-luciferase reporter gene assay. LncRNA function as competitive endogenous RNA (ceRNA) in keloid was further verified by in keloid fibroblasts (KFs) and in nude mice with subcutaneous keloids.

Results: BTF3 expression was up-regulated in keloid tissues. The targeting relationship between BTF3 and miR-15b-5p was confirmed by dual-luciferase reporter gene assay. miR-15b-5p overexpression inhibited BTF3, Bcl-2, Cyclin D1, C-myc, Collagen I, MMP2, MMP9, N-cadherin, and ZEB2 expressions in KFs, inhibited cell proliferation and migration, while promoted E-cadherin levels. BTF3 overexpression reversed miR-15b-5p effects on KFs. Bioinformatics analysis as well as clinical and cellular experiments confirmed that the lncRNA ZNF252P-AS1 was highly expressed in keloid/KFs. Dual-luciferase reporter gene assays confirmed the targeting relationship between lncRNA ZNF252P-AS1 and miR-15b-5p. LncRNA ZNF252P-AS1 overexpression inhibited miR-15b-5p and E-cadherin levels, upregulated BTF3, Bcl-2, Cyclin D1, C-myc, Collagen I, MMP2, MMP9, N-cadherin, and ZEB2 expressions, increased cell proliferation and migration, and activated JAK2/STAT3 pathway, while miR-15b-5p overexpression reversed this effect. The in vivo results were consistent with in vitro results. In vivo experiments further confirmed that lncRNA ZNF252P-AS1 reduced keloid volume and weight.

Conclusion: lncRNA ZNF252P-AS1 is a potential target for keloid treatment.

Keywords: BTF3; Fibroblast; Keloid; LncRNA-ZNF252P-AS1; MiR-15b-5p; STAT3.

MeSH terms

Animals
Cadherins / genetics
Cell Line, Tumor
Cell Movement / genetics
Cell Proliferation / genetics
Collagen / metabolism
Cyclin D1 / genetics
Fibroblasts / metabolism
Gene Expression Regulation, Neoplastic
Humans
Keloid* / genetics
Matrix Metalloproteinase 2 / metabolism
Matrix Metalloproteinase 9 / genetics
Matrix Metalloproteinase 9 / metabolism
Mice
Mice, Nude
MicroRNAs* / genetics
MicroRNAs* / metabolism
RNA, Long Noncoding* / genetics
RNA, Long Noncoding* / metabolism
Signal Transduction / genetics

Substances

Cadherins
Collagen
Cyclin D1
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
MicroRNAs
RNA, Long Noncoding
Stat3 protein, mouse
transcription factor BTF3
MIRN15 microRNA, human