DKK3 regulates cell proliferation, apoptosis and collagen synthesis in keloid fibroblasts via TGF-β1/Smad signaling pathway

Li, Yang; Liu, Hengxin; Liang, Yingzi; Peng, Pai; Ma, Xianjie; Zhang, Xi

doi:10.1016/j.biopha.2017.03.044

Cited by 43 publications

(40 citation statements)

References 30 publications

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“…Moreover, analysis of DKK3 gene expression by q-RT-PCR indicated that decitabine treatment increased DKK3 mRNA levels, both in PC3 cells, as previously reported [ 39 ], and in C4-2B cells ( Figure 1 B). Given the links between Dkk-3 and TGF-β-signaling [ 20 , 24 , 40 ], gene reporter assays were carried out to determine the effects of decitabine on TGF-β-dependent transcription using pGL3-CAGA12, which encodes the luciferase gene fused to 12 repeats of a Smad-binding site, and renilla, to control for transfection efficiency. Decitabine reduced TGF-β-dependent gene reporter activity in PC3 cells but not in C4-2B cells ( Figure 1 C), consistent with Dkk-3 inhibition of TGF-β signaling in PC3 cells and not in C4-2B cells, which do not express TGFBR2 [ 41 ].…”

Section: Resultsmentioning

confidence: 99%

CRISPR-Mediated Reactivation of DKK3 Expression Attenuates TGF-β Signaling in Prostate Cancer

Kardooni

González-Gualda

Stylianakis

et al. 2018

Cancers

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The DKK3 gene encodes a secreted protein, Dkk-3, that inhibits prostate tumor growth and metastasis. DKK3 is downregulated by promoter methylation in many types of cancer, including prostate cancer. Gene silencing studies have shown that Dkk-3 maintains normal prostate epithelial cell homeostasis by limiting TGF-β/Smad signaling. While ectopic expression of Dkk-3 leads to prostate cancer cell apoptosis, it is unclear if Dkk-3 has a physiological role in cancer cells. Here, we show that treatment of PC3 prostate cancer cells with the DNA methyltransferase (DNMT) inhibitor decitabine demethylates the DKK3 promoter, induces DKK3 expression, and inhibits TGF-β/Smad-dependent transcriptional activity. Direct induction of DKK3 expression using CRISPR-dCas9-VPR also inhibited TGF-β/Smad-dependent transcription and attenuated PC3 cell migration and proliferation. These effects were not observed in C4-2B cells, which do not respond to TGF-β. TGF-β signals can regulate gene expression directly via SMAD proteins and indirectly by increasing DNMT expression, leading to promoter methylation. Analysis of genes downregulated by promoter methylation and predicted to be regulated by TGF-β found that DKK3 induction increased expression of PTGS2, which encodes cyclooxygenase-2. Together, these observations provide support for using CRISPR-mediated induction of DKK3 as a potential therapeutic approach for prostate cancer and highlight complexities in Dkk-3 regulation of TGF-β signaling.

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Section: Resultsmentioning

confidence: 99%

CRISPR-Mediated Reactivation of DKK3 Expression Attenuates TGF-β Signaling in Prostate Cancer

Kardooni

González-Gualda

Stylianakis

et al. 2018

Cancers

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“…Transforming growth factor β1 (TGF-β1), ‘master cytokine’ in fibrosis ( 5 ), plays a crucial role in the pathogenesis of intestinal fibrosis, including fibroblasts proliferation and differentiation into myofibroblasts, and ECM production ( 6 , 7 ). Therefore, TGF-β1-stimulated fibroblasts have been regarded as a good cellular model for evaluating the efficacy of antifibrotic agents ( 8 , 9 ). It has been reported that induction of apoptosis and inhibition of proliferation and profibrogenic effects in fibroblasts have shown potential in the treatment of intestinal fibrosis ( 10 ).…”

Section: Introductionmentioning

confidence: 99%

Pirfenidone suppresses TGF‑β1‑induced human intestinal fibroblasts activities by regulating proliferation and apoptosis via the inhibition of the Smad and PI3K/AKT signaling pathway

2018

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Intestinal fibroblasts, the main effector cells of intestinal fibrosis, are considered to be a good target for anti-fibrotic therapy. The aim of the present study was to examine the effects of pirfenidone (PFD) on human intestinal fibroblasts (HIFs) stimulated by transforming growth factor (TGF)-β1 and to explore the potential mechanism. Prior to stimulation with TGF-β1 (10 ng/ml), HIFs were treated with or without PFD (1 mg/ml). Cell proliferation was determined by Cell Counting Kit (CCK)-8 and colony formation assays, and cell apoptosis was assessed using flow cytometry and a TUNEL assay. Reverse transcription-quantitative polymerase chain reaction and western blotting were performed to evaluate the mRNA and protein expressions of α-smooth muscle actin (α-SMA), collagen I and fibronectin. The protein expression of TGF-β1/mothers against decapentaplegic homolog (Smad) and phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathways was evaluated by western blotting. CCK-8 and colony formation assays demonstrated that PFD significantly inhibited cell proliferation in HIFs stimulated with TGF-β1. Flow cytometry and TUNEL assays revealed that PFD treatment significantly enhanced apoptosis in TGF-β1-stimulated HIFs. In addition, PFD markedly reduced TGF-β1-induced HIF activities, such as myofibroblast differentiation (α-SMA), and collagen production (collagen I and fibronectin). These effects of PFD were mediated by the inhibition of the TGF-β1/Smad and PI3K/AKT signaling pathways. Therefore, the present study demonstrated that PFD reduced TGF-β1-induced fibrogenic activities of HIFs, suggesting that PFD may be a potential therapeutic agent for intestinal fibrosis.

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“…However, DKK3 is the most structurally divergent member of the Dickkopf family (Krupnik et al, 1999) and has varied effects on Wnt, ranging from no effect (Krupnik et al, 1999;Pinho Christof Niehrs, 2007;Romero et al, 2013) to promoting (Ferrari et al, 2019;Nakamura and Hackam, 2010;Yin et al, 2018) or inhibiting Wnt (Bhattacharyya et al, 2017;Leonard et al, 2017;Liu et al, 2019;Sharma Das et al, 2013). DKK3 also functions as both a positive and negative regulator of TGFβ signaling, depending on the model (Al Shareef et al, 2018;Busceti et al, 2017;Kardooni et al, 2018;Li et al, 2017;Pinho Christof Niehrs, 2007;Romero et al, 2013;Wang Z et al, 2015), but DKK3 did not impact TGFβ in this study (data not shown). Despite conflicting reports of DKK3's signaling targets, it has consistently been shown to restrain cell proliferation (Kawano et al, 2006;Leonard et al, 2017).…”

Section: 25d Regulates Dkk3 In Lineage-committed Cellsmentioning

confidence: 65%

Vitamin D Sufficiency Enhances Differentiation of Patient-Derived Prostate Epithelial Organoids

McCray

Pacheco

Baumann

et al. 2020

Preprint

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Vitamin D is an essential steroid hormone that regulates systemic calcium homeostasis and cell fate decisions, including differentiation in many cell types. The prostate gland is hormonally regulated, requiring steroids for proliferation and terminal differentiation of secretory luminal cells. Vitamin D deficiency is associated with higher risk of lethal prostate cancer with an aggressive dedifferentiated pathology, linking vitamin D sufficiency to epithelial differentiation homeostasis. To determine regulation of prostate epithelial differentiation by vitamin D status, patient-derived benign prostate epithelial organoids were maintained in vitamin D deficient (vehicle) or sufficient (10 nM 1,25-dihydroxyvitamin D) conditions and assessed by phenotype and single cell RNA sequencing. Mechanistic validation demonstrated that vitamin D sufficiency promoted organoid growth and accelerated differentiation of lineage-committed cells by inhibiting canonical Wnt activity and Wnt family member DKK3. Wnt dysregulation is a known contributor to aggressive prostate cancer, thus these findings further link vitamin D deficiency to lethal disease.

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DKK3 regulates cell proliferation, apoptosis and collagen synthesis in keloid fibroblasts via TGF-β1/Smad signaling pathway

Cited by 43 publications

References 30 publications

CRISPR-Mediated Reactivation of DKK3 Expression Attenuates TGF-β Signaling in Prostate Cancer

CRISPR-Mediated Reactivation of DKK3 Expression Attenuates TGF-β Signaling in Prostate Cancer

Pirfenidone suppresses TGF‑β1‑induced human intestinal fibroblasts activities by regulating proliferation and apoptosis via the inhibition of the Smad and PI3K/AKT signaling pathway

Vitamin D Sufficiency Enhances Differentiation of Patient-Derived Prostate Epithelial Organoids

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