Hsa_circ_0001445 regulates acne inflammation via sponging miR-1298-5p targeting ESR1

Abstract

Introduction: Acne is a prevalent inflammatory skin condition that occurs in adolescents and can persist into adulthood. CircRNA has been reported to be widely involved in a variety of human diseases. However, the regulatory mechanism of hsa_circ_0001445 in acne has rarely been reported. The purpose is to research the role of hsa_circ_0001445 in acne-induced inflammation and its molecular regulatory mechanism to provide a foundation for the exploration of acne-targeting drugs.

Material and methods: We used the GEO, starBase, and GeneCards databases for bioinformatics analysis. The binding sequences of miR-1298-5p and hsa_circ_0001445 or ESR1 mRNA were predicted by the Circular RNA Interactome or starBase database. Double luciferase reporting assay was applied to verify the regulatory relationship between hsa_circ_0001445, miR-1298-5p, and ESR1. RT-qPCR was used to detect levels of hsa_circ_0001445, miR-1298-5p, and ESR1 mRNA. The secretion levels of interleukin (IL)-6, IL-8, and tumor necrosis factor α (TNF-α) were measured using an ELISA kit.

Results: The luciferase activity was weakened by miR-1298-5p mimics in human keratinocytes and sebocytes transfected with wild-type (wt)-circ1445 and wt-ESR1, respectively. Moreover, the overexpression of hsa_circ_0001445 reduced the miR-1298-5p level and reversed the elevation of IL-6, IL-8, and TNF-α levels in Bio-C. acnes-stimulated keratinocytes and sebocytes. In turn, transfection of miR-1298-5p mimics partially eliminated the inflammatory inhibition of hsa_circ_0001445, which was reversed by co-transfection of pcDNA-ESR1.

Conclusions: Hsa_circ_0001445 improved acne inflammation via sponging miR-1298-5p targeting ESR1.

Keywords: ESR1; acne; hsa_circ_0001445; inflammation; miR-1298-5p.

Fig. 1

Hsa_circ_0001445 showed low expression in acne. A) The expression graph of hsa_circ_0001445 in acne patients and healthy volunteers from the GSE212605 dataset. B, C) The expression of hsa_circ_0001445 in normal or Bio-C. acnes-stimulated human keratinocytes and sebocytes (n = 4). ***p < 0.001. n – number of experiments

Fig. 2

Hsa_circ_0001445 inhibits acne inflammation. A) Hsa_circ_0001445 level in Bio-C. acnes-stimulated human keratinocytes and sebocytes transfected with hsa_circ_0001445 overexpression vector (n = 4). B-D) Levels of IL-6, IL-8, and TNF-α in Bio-C. acnes-stimulated human keratinocytes and sebocytes transfected with or without hsa_circ_0001445 overexpression vector (n = 4). ***p < 0.001 (vs. control), ###p < 0.001 (vs. Bio-C. acnes + ov-ctrl). n – number of experiments

Fig. 3

Hsa_circ_0001445 sponges miR-1298-5p. A) Venn diagram of high-expressed miRNAs in acne patients from GSE212605 dataset, as well as the target miRNAs of hsa_circ_0001445. B) Expression graph of miR-5094, miR-127-5p, and miR-1298-5p in acne patients and healthy volunteers from GSE212605 dataset. C, D) Expression of miR-1298-5p in normal or Bio-C. acnes-stimulated human keratinocytes and sebocytes (n = 4). E) Binding sequence of hsa_circ_0001445 and miR-1298-5p predicted using Circular RNA Interactome database. ***p < 0.001. n – number of experiments F, G) Effects of miR-1298-5p mimics on the luciferase activity of keratinocytes and sebocytes transfected with WT-circ1445 or MUT-circ1445 vectors (n = 4). ***p < 0.001. n – number of experiments

Fig. 3

Hsa_circ_0001445 sponges miR-1298-5p. A) Venn diagram of high-expressed miRNAs in acne patients from GSE212605 dataset, as well as the target miRNAs of hsa_circ_0001445. B) Expression graph of miR-5094, miR-127-5p, and miR-1298-5p in acne patients and healthy volunteers from GSE212605 dataset. C, D) Expression of miR-1298-5p in normal or Bio-C. acnes-stimulated human keratinocytes and sebocytes (n = 4). E) Binding sequence of hsa_circ_0001445 and miR-1298-5p predicted using Circular RNA Interactome database. ***p < 0.001. n – number of experiments F, G) Effects of miR-1298-5p mimics on the luciferase activity of keratinocytes and sebocytes transfected with WT-circ1445 or MUT-circ1445 vectors (n = 4). ***p < 0.001. n – number of experiments

Fig. 4

Hsa_circ_0001445 alleviates acne inflammation by targeting and negatively regulating miR-1298-5p. A) Effects of hsa_circ_0001445 overexpression or in combination with miR-1298-5p mimics on the hsa_circ_0001445 level of Bio-C. acnes-stimulated keratinocytes and sebocytes (n = 4). B-D) Effects of hsa_circ_0001445 overexpression or in combination with miR-1298-5p mimics on the levels of IL-6, IL-8, and TNF-α in Bio-C. acnes-stimulated keratinocytes and sebocytes (n = 4). **p < 0.01, ***p < 0.001 (vs. Bio-C. acnes + ov-ctrl), ##p < 0.01, ###p < 0.001 (vs. Bio-C. acnes + ov-circ1445 + mimNC). n – number of experiments

Fig. 5

MiR-1298-5p directly targets and negatively regulates ESR1. A) Binding sequence of miR-1298-5p and ESR1 mRNA predicted using starBase database. B, C) Effects of miR-1298-5p mimics on luciferase activity of keratinocytes and sebocytes transfected with WT-ESR1 or MUT-ESR1 vectors (n = 4). D) Effects of miR-1298-5p mimics alone or in combination with pcDNA-ESR1 on the mRNA level of ESR1 in Bio-C. acnes-stimulated keratinocytes and sebocytes after hsa_circ_0001445 overexpression (n = 4). E, F) Effects of miR-1298-5p mimics alone or in combination with pcDNA-ESR1 on IL-6, IL-8, and TNF-α levels in Bio-C. acnes-stimulated keratinocytes and sebocytes after hsa_circ_0001445 overexpression (n = 4). **p < 0.01, ***p < 0.001 (vs. Bio-C. acnes + ov-circ1445 + mimNC). ###p < 0.001 (vs. Bio-C. acnes + ov-circ1445 + mim1298-5p + pcDNA). n – number of experiments