PAX5 activates telomerase activity and proliferation in keloid fibroblasts by transcriptional regulation of SND1, thus promoting keloid growth in burn-injured skin

Abstract

Objective

Staphylococcal nuclease domain-containing 1 (SND1) that functioned as an oncogene in a variety of tumors was upregulated in burn-injured skin tissues, and this study aims to investigate the effect of SND1 on keloid and elucidate the underlying mechanism.

Methods

Keloid fibroblasts (KFs) and normal skin fibroblasts (NFs) were isolated from the keloid tissues and adjacent normal skin tissues of keloid patients. The SND1 expression was assessed in keloid tissues and KFs with Western blot assay. Gain- and loss-of-function experiments were performed to investigate the role of SND1 in proliferation, colony formation, telomerase activity, expression of fibrogenic genes and production of pro-inflammatory factors in KFs. Chromatin immunoprecipitation (CHIP) and Dual-luciferase reporter gene assays were used to verify the interaction of Paired-box gene 5 (PAX5) on SND1 promoter. Then, a series of rescue experiments were performed to verify the effects of SND1 overexpression on PAX5 knockdown-mediated KF functions. Finally, the role of SND1 in keloid formation in vivo was validated in mice with keloid implantation.

Results

SND1 was upregulated in keloid tissues and KFs. SND1 positively regulated proliferation, colony formation, telomerase activity, production of pro-inflammatory factors and expression of fibrogenic genes. PAX5 directly bound to the SND1 promoter to transcriptionally regulate SND1 expression and positively regulated SND1-mediated KF functions via the ERK/JNK pathway. In vivo assay further demonstrated that SND1 displayed a positive effect on keloid formation.

Conclusion

SND1 transcriptionally regulated by PAX5 promotes keloid formation through activating telomerase activity via the ERK/JNK signaling pathways, which provides a promising therapeutic target for clinical treatment of burned skin keloid.

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Abbreviations

SND1:

Staphylococcal nuclease domain-containing 1

PAX5:

Paired-box gene 5

CHIP:

Chromatin immunoprecipitation

KFs:

Keloid fibroblasts

NFs:

Normal fibroblasts

CCK-8:

Cell Counting Kit-8

RT-qPCR:

Quantitative real-time polymerase chain reaction

TRAP:

Telomeric repeat amplification protocol

ANOVA:

One-way analysis of variance

MAPK:

Mitogen-activated protein kinase

ERK:

Extracellular signal-regulated kinase

JNK:

C-Jun N-terminal kinase

FBS:

Fetal bovine serum

ELISA:

Enzyme linked immunosorbent assay

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Correspondence to Yong Song.

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Qin, G., Sun, Y., Guo, Y. et al. PAX5 activates telomerase activity and proliferation in keloid fibroblasts by transcriptional regulation of SND1, thus promoting keloid growth in burn-injured skin. Inflamm. Res. (2021). https://doi.org/10.1007/s00011-021-01444-3

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Keywords

  • Keloid
  • SND1
  • PAX5
  • Telomerase activity
  • Burn
  • Transcriptional regulation