Indirubin inhibits Wnt/β-catenin signal pathway via promoter demethylation of WIF-1
| Autor: | Yong Feng Chen, Shou Gang Liu, Guang Pu Luo, Yong Bin Qu |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
DNA (Cytosine-5-)-Methyltransferase 1
Indoles Wnt signal pathway Wif-1 Apoptosis Filaggrin Proteins chemistry.chemical_compound HaCaT Cells Humans Psoriasis Viability assay Involucrin Wnt Signaling Pathway beta Catenin Adaptor Proteins Signal Transducing Cell Proliferation Indirubin integumentary system Chemistry DNMT1 Wnt signaling pathway lcsh:Other systems of medicine Cell Cycle Checkpoints Cell cycle lcsh:RZ201-999 Molecular biology HaCaT Complementary and alternative medicine Catenin Loricrin Research Article |
| Zdroj: | BMC Complementary Medicine and Therapies BMC Complementary Medicine and Therapies, Vol 20, Iss 1, Pp 1-10 (2020) |
| ISSN: | 2662-7671 |
| Popis: | BackgroundPsoriasis 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.MethodsThe 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.ResultsIndirubin 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.ConclusionOur 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|