Top

BMC Complementary Medicine and Therapies

Published in:

01-12-2020 | Vulgar Psoriasis | Research article

Indirubin inhibits Wnt/β-catenin signal pathway via promoter demethylation of WIF-1

Authors: Shou Gang Liu, Guang Pu Luo, Yong Bin Qu, Yong Feng Chen

Published in: BMC Complementary Medicine and Therapies | Issue 1/2020

Abstract

Background

Psoriasis is a common inflammatory skin disease. Abnormal proliferation of keratinocytes is one of the psoriatic histopathological features. Indirubin has an essential effect on the proliferation and activation of keratinocytes; however, in psoriasis, the specific mechanism of action of indirubin on keratinocytes is unclear. In the present study, we revealed the effects of indirubin on DNA methyltransferase 1 (DNMT1), wnt inhibitory factor 1 (wif-1), and wnt/β-catenin signal pathway, in the meantime, we explored the effects of indirubin on proliferation, cell cycle and the apoptosis of HaCaT cells.

Methods

The expression of DNMT1, wif-1, Frizzled2, Frizzled5, and β-catenin in HaCaT cells treated with different concentrations of indirubin were detected by Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR). The expression levels of DNMT1 and wif-1 were observed after treated with different concentrations of indirubin by enzyme-linked immunosorbent assay (ELISA). The wif-1 promoter methylation status was detected by DNA methylation-specific PCR (MSP). The transcriptional activities of wif-1 and β-catenin were discovered by a luciferase reporter gene system. Cell viability was determined by Cell Counting Kit-8 (CCK8) method. The cell cycle was detected by flow cytometry. The apoptotic cells were surveyed by the apoptosis kit. The expression of Inolucrin, Loricrin, Filaggrin, Keratin 17, and transcriptional activation of transglutaminase 1(TGase1) were detected by Western blotting.

Results

Indirubin inhibited the expression of DNMT1 and the methylation of the wif-1 promoter. In the wnt signal pathway, indirubin restored the protein expression of wif-1 and inhibited expression of Frizzled2, Frizzled5, and β-catenin. Besides, indirubin inhibited the proliferation of HaCaT cells, induced apoptosis, and arrest cell cycle. We also reported that indirubin could down-regulate the expression of Involucrin, TGase 1, and keratin 17, but the expression of Filaggrin and Loricrin had no significant effect.

Conclusion

Our research showed that indirubin promoted the demethylation of wif-1 and suppressed the wnt/β-catenin signal pathway, thereby exerted an anti-proliferative effect. This study reveals the anti-proliferation mechanism of indirubin, which may provide an effective option for the treatment of proliferative diseases.

Rendon A, Schäkel K. Psoriasis pathogenesis and treatment. Int J Mol Sci. 2019;20(6):1475.PubMedCentral

Boehncke, W. H, Schön, M. P, Psoriasis Lancet2015, 386 (9997): 983–994.

Reuter J, Wölfle U, Weckesser S, Schempp C. Which Plant for Which Skin Disease? Part 1: atopic dermatitis, psoriasis, acne, Condyloma and herpes simplex. J Dtsch Dermatol Ges. 2010;8(10):788–96.PubMed

Huang M, Wang L, Zeng S, Qiu Q, Zou Y, Shi M, Xu H, Liang L. Indirubin inhibits the migration, invasion, and activation of fibroblast-like synoviocytes from rheumatoid arthritis patients. Inflamm Res. 2017;66(5):433–40.PubMed

Hsieh WL, Lin YK, Tsai CN, Wang TM, Chen TY. JongHwei S PangIndirubin, an acting component of indigo naturalis, inhibits EGFR activation and EGF-induced CDC25B gene expression in epidermal keratinocytes. J Dermatol Sci. 2012;67(2):140–6.PubMed

Xie X, Di T, Wang Y, Wang M, Meng Y, Lin Y, Xu X, Li P, Zhao J. Indirubin ameliorates imiquimod-induced psoriasis-like skin lesions in mice by inhibiting inflammatory responses mediated by IL-17A-producing γδ T cells. Mol Immunol. 2018;101:386–95.PubMed

Lin Y-K, See L-C, Huang Y-H, Chi C-C, C-Y Hui R. Comparison of indirubin concentrations in indigo naturalis ointment for psoriasis treatment: a randomized, double-blind, dosage-controlled trial. Br J Dermatol. 2018;178(1):124–31.PubMed

Wenming Wang, Xiaoling Yu, Chao Wu, and Hongzhong Jin.IL-36γ inhibits differentiation and induces inflammation of keratinocyte via Wnt signaling pathway in psoriasis.Int J Med Sci. 2017; 14(10):1002–1007.

Zhang Y, Chen T, Zhang D, Zheng Y, Peng Z, Feng Y, Xiao S, Li Z. Wnt/β-catenin and Wnt5a/Ca pathways regulate proliferation and apoptosis of keratinocytes in psoriasis lesions. Cell Physiol Biochem. 2015;36(5):1890–902.PubMed

10.

Gudjonsson JE, Johnston A, Stoll SW, Riblett MB, Xing X, Kochkodan JJ, Ding J, Nair RP, Aphale A, Voorhees JJ, Elder JT. Evidence for altered Wnt signaling in psoriatic skin. J Invest Dermatol. 2010;130(7):1849–59.PubMedPubMedCentral

11.

Deng Y, Yu B, Cheng Q, Jin J, You H, Ke R, NingTang QS, Shu H, Yao G, Zhang Z, Qin W. Epigenetic silencing of WIF- in hepatocellular carcinomas. J Cancer Res Clin Oncol. 2010;136(8):1161–7.PubMed

12.

Reguart N, He B, Xu Z, You L, Lee AY, Mazieres J, Mikami I, Batra S, Rosell R, McCormick F, Jablons DM. Cloning and characterization of the promoter of human Wnt inhibitory factor-1. Biochem Biophys Res Commun. 2004;323(1):229–34.PubMed

13.

Lisa D Moore, Thuc Le, Guoping Fan. DNA methylation and its basic function. Neuropsychopharmacology. 2013;38(1):23–38.

14.

Chen M, Chen Z-Q, Cui P-G, Yao X, Li Y-M, Li A-S, Gong J-Q, Cao Y-H. The methylation pattern of p16INK4a gene promoter in psoriatic epidermis and its clinical significance. Br J Dermatol. 2008;158(5):987–93.PubMed

15.

Rountree MR, Bachman KE, Herman JG, Baylin SB. DNAmethylation, chromatin, inheritance, and cancer. Oncogene.2001; 20 (24): 3156–3165.

16.

Zhang P, Yuwen S, Chen H, Zhao M, Lu Q. Abnormal DNA methylation in skin lesions and PBMCs of patients with psoriasis vulgaris. J Dermatol Sci. 2010;60(1):40–2.PubMed

17.

Anastas JN, Moon RT. WNT signalling pathways as therapeutic targets in cancer. Nat Rev Cancer. 2013;13(1):11–26.PubMed

18.

Yu X, Yan N, Li Z, Hua Y, Chen W. FGF19 sustains the high proliferative ability of keratinocytes in psoriasis through the regulation of Wnt/GSK-3β/β-catenin signalling via FGFR4. Clin Exp Pharmacol Physiol. 2019;46(8):761–9.PubMed

19.

Lin B, Hong HJ, Jiang XJ, Li CZ, Zhu SY, Tang NH, Wang XQ, She FF, Chen YL. WNT inhibitory factor 1 promoter hypermethylation is an early event during gallbladder cancer tumorigenesis that predicts poor survival. Gene. 2017;622:42–9.PubMed

20.

Xue X, Wu J, Li J, Xu J, Dai H, Tao C, Li C, Jinhong H. Indirubin attenuates mouse psoriasis-like skin lesion in a CD274-dependent manner: an achievement of RNA sequencing. Biosci Rep. 2018;38(6):BSR20180958.PubMedPubMedCentral

21.

Tan M, Wu J, Cai Y. Suppression of Wnt signaling by the miR-29 family is mediated by demethylation of WIF- in non-small-cell lung cancer. Biochem Biophys Res Commun. 2013;438(4):673–9.PubMed

22.

Uluçkan Ö, Jimenez M, Karbach S, Jeschke A, Graña O, Keller J, Busse B, Croxford AL, Finzel S, Koenders M, van den Berg W, Schinke T, Amling M, Waisman A, Schett G, Wagner EF. Chronic skin inflammation leads to bone loss by IL-17-mediated inhibition of Wnt signaling in osteoblasts. Sci Transl Med. 2016;8(330):330–7.

23.

Schroeder WT, Thacher SM, Stewart-Galetka S, Annarella M, Chema D, Siciliano MJ, Davies PJ, Tang HY, Sowa BA, Duvic M. Type I keratinocyte transglutaminase: expression in human skin and psoriasis. J Invest Dermatol. 1992;99:27–34.PubMed

24.

Richard L Eckert , Michael T Sturniolo, Ann-Marie Broome, Monica Ruse, Ellen A Rorke. Transglutaminase function in epidermis. J Invest Dermatol.2005; 124 (3):481–492.

25.

Rice RH, Green H. The cornified envelope of terminally differentiated human epidermal keratinocytes consists of cross-linked protein. Cell. 1977;11(2):417–22.PubMed

26.

Hampton PJ, Ross OK, Reynolds NJ. Increased nuclear beta-catenin in suprabasal involved psoriatic epidermis. Br J Dermatol. 2007;157(6):1168–77.PubMed

27.

Depianto D, Kerns ML, Dlugosz AA, Coulombe PA. Keratin 17 promotes epithelial proliferation and tumor growth by polarizing the immune response in skin. Nat Genet. 2010;42(10):910–4.PubMedPubMedCentral

28.

Chung BM, Arutyunov A, Ilagan E, Yao N, Wills-Karp M. Pierre A CoulombeRegulation of C-X-C chemokine gene expression by keratin 17 and hnRNP K in skin tumor keratinocytes. J Cell Biol. 2015;208(5):613–27.PubMedPubMedCentral

29.

Liang J, Wang G. Keratin 17: a critical player in the pathogenesis of psoriasis. Med Res Rev. 2014;34(2):438–54.

30.

Iizuka H, Takahashi H, Honma M, Ishida-Yamamoto A. Unique keratinization process in psoriasis: late differentiation markers are abolished because of the premature cell death. J Dermatol. 2004;31(4):271–6.PubMed

Title: Indirubin inhibits Wnt/β-catenin signal pathway via promoter demethylation of WIF-1
Authors: Shou Gang Liu
Guang Pu Luo
Yong Bin Qu
Yong Feng Chen
Publication date: 01-12-2020
Publisher: BioMed Central
Keyword: Vulgar Psoriasis
Published in: BMC Complementary Medicine and Therapies / Issue 1/2020
Electronic ISSN: 2662-7671
DOI: https://doi.org/10.1186/s12906-020-03045-9

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Indirubin inhibits Wnt/β-catenin signal pathway via promoter demethylation of WIF-1

Abstract

Background

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2020

Comparative in vitro analysis of inhibition of rhinovirus and influenza virus replication by mucoactive secretolytic agents and plant extracts

Anti-inflammatory activity of 3-cinnamoyltribuloside and its metabolomic analysis in LPS-activated RAW 264.7 cells

Curcumin inhibits the migration of osteoclast precursors and osteoclastogenesis by repressing CCL3 production

Parents’ experiences of information-seeking and decision-making regarding complementary medicine for children with autism spectrum disorder: a qualitative study

The effect of Malus doumeri leaf flavonoids on oxidative stress injury induced by hydrogen peroxide (H2O2) in human embryonic kidney 293 T cells

Comparison of the acute toxicity, analgesic and anti-inflammatory activities and chemical composition changes in Rhizoma anemones Raddeanae caused by vinegar processing