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Knockdown of lncRNA-ATB suppresses autocrine secretion of TGF-β2 by targeting ZNF217 via miR-200c in keloid fibroblasts.

Zhu HY, Bai WD, Li C, Zheng Z, Guan H, Liu JQ, Yang XK, Han SC, Gao JX, Wang HT, Hu DH - Sci Rep (2016)

Bottom Line: Through gain- and loss-of-function studies, we demonstrated that knockdown of lncRNA-ATB decreased autocrine secretion of TGF-β2 and ZNF217 expression but upregulated expression of miR-200c in KFs.Stable downregulation of ZNF217 expression decreased the autocrine secretion of TGF-β2. miR-200c was endogenously associated with lncRNA-ATB, and inhibition of miR-200c overcame the decrease in ZNF217 expression in KFs.Taken together, these findings indicate that lncRNA-ATB governs the autocrine secretion of TGF-β2 in KFs, at least in part, by downregulating the expression level of ZNF217 via miR-200c, suggesting a signaling axis consisting of lncRNA-ATB/miR-200c/ZNF217/TGF-β2.

View Article: PubMed Central - PubMed

Affiliation: Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, Shaanxi, People's Republic of China.

ABSTRACT
Abnormally high activation of transforming growth factor-β (TGF-β) signaling has been demonstrated to be involved in the initiation and progression of keloids. However, the functional role of long non-coding RNA (lncRNA)-activated by TGF-β (lncRNA-ATB) in keloids has not been documented. Here we investigated the role of lncRNA-ATB in the autocrine secretion of TGF-β in keloid fibroblasts (KFs) and explored the underlying molecular mechanism. Using immunohistochemistry and quantitative RT-PCR analysis, we showed that lncRNA-ATB and ZNF217, a transcriptional activator of TGF-β, were overexpressed and miR-200c, which targets ZNF217, was under-expressed in keloid tissue and keloid fibroblasts. Through gain- and loss-of-function studies, we demonstrated that knockdown of lncRNA-ATB decreased autocrine secretion of TGF-β2 and ZNF217 expression but upregulated expression of miR-200c in KFs. Stable downregulation of ZNF217 expression decreased the autocrine secretion of TGF-β2. miR-200c was endogenously associated with lncRNA-ATB, and inhibition of miR-200c overcame the decrease in ZNF217 expression in KFs. Taken together, these findings indicate that lncRNA-ATB governs the autocrine secretion of TGF-β2 in KFs, at least in part, by downregulating the expression level of ZNF217 via miR-200c, suggesting a signaling axis consisting of lncRNA-ATB/miR-200c/ZNF217/TGF-β2. These findings may provide potential biomarkers and targets for novel diagnostic and therapeutic approaches for keloids.

No MeSH data available.


Related in: MedlinePlus

ZNF217 expression in keloid tissue and keloid fibroblasts.(A) Immunohistochemical staining for ZNF217 in keloid (K) tissues and paired normal skin (NS) tissues. Upper, NS; lower, K; left, ×40; right, ×200; (B) Comparison between K and NS tissues regarding ZNF217 expression in terms of mean intensity; (C) Western blot analysis of ZNF217 protein expression in normal fibroblasts (NF) and keloid fibroblasts (KF), with β-actin being used as an internal control; (D) qRT-PCR determination of relative expression levels of ZNF217 in NF and KF.
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f2: ZNF217 expression in keloid tissue and keloid fibroblasts.(A) Immunohistochemical staining for ZNF217 in keloid (K) tissues and paired normal skin (NS) tissues. Upper, NS; lower, K; left, ×40; right, ×200; (B) Comparison between K and NS tissues regarding ZNF217 expression in terms of mean intensity; (C) Western blot analysis of ZNF217 protein expression in normal fibroblasts (NF) and keloid fibroblasts (KF), with β-actin being used as an internal control; (D) qRT-PCR determination of relative expression levels of ZNF217 in NF and KF.

Mentions: Given that ZNF217, a transcriptional activator of TGF-β, promotes EMT in breast cancer8 and that TGF-β signaling is elevated in keloids, we surmised that ZNF217 might modulate the autocrine secretion of TGF-β2 in keloids. We first determined the expression of ZNF217 protein by immunohistochemistry in keloid tissue samples from our retrospective cohort of 57 patients with keloids, whose clinicopathological data are presented in Supplemental Table 1. By signal intensity analysis, we found that ZNF217 expression was significantly increased in keloid tissue compared with normal skin tissue (P = 0.013; Fig. 2A,B). To confirm this finding, we next evaluated ZNF217 protein and mRNA expression in keloid tissue and normal skin tissue by Western blotting and qRT-PCR, respectively. The results showed that ZNF217 was overexpressed in keloid fibroblasts at both the protein and mRNA levels, compared to levels in normal skin tissue (Fig. 2C,D). Altogether, these data suggest that ZNF217 is overexpressed in keloid tissue and fibroblasts.


Knockdown of lncRNA-ATB suppresses autocrine secretion of TGF-β2 by targeting ZNF217 via miR-200c in keloid fibroblasts.

Zhu HY, Bai WD, Li C, Zheng Z, Guan H, Liu JQ, Yang XK, Han SC, Gao JX, Wang HT, Hu DH - Sci Rep (2016)

ZNF217 expression in keloid tissue and keloid fibroblasts.(A) Immunohistochemical staining for ZNF217 in keloid (K) tissues and paired normal skin (NS) tissues. Upper, NS; lower, K; left, ×40; right, ×200; (B) Comparison between K and NS tissues regarding ZNF217 expression in terms of mean intensity; (C) Western blot analysis of ZNF217 protein expression in normal fibroblasts (NF) and keloid fibroblasts (KF), with β-actin being used as an internal control; (D) qRT-PCR determination of relative expression levels of ZNF217 in NF and KF.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4835760&req=5

f2: ZNF217 expression in keloid tissue and keloid fibroblasts.(A) Immunohistochemical staining for ZNF217 in keloid (K) tissues and paired normal skin (NS) tissues. Upper, NS; lower, K; left, ×40; right, ×200; (B) Comparison between K and NS tissues regarding ZNF217 expression in terms of mean intensity; (C) Western blot analysis of ZNF217 protein expression in normal fibroblasts (NF) and keloid fibroblasts (KF), with β-actin being used as an internal control; (D) qRT-PCR determination of relative expression levels of ZNF217 in NF and KF.
Mentions: Given that ZNF217, a transcriptional activator of TGF-β, promotes EMT in breast cancer8 and that TGF-β signaling is elevated in keloids, we surmised that ZNF217 might modulate the autocrine secretion of TGF-β2 in keloids. We first determined the expression of ZNF217 protein by immunohistochemistry in keloid tissue samples from our retrospective cohort of 57 patients with keloids, whose clinicopathological data are presented in Supplemental Table 1. By signal intensity analysis, we found that ZNF217 expression was significantly increased in keloid tissue compared with normal skin tissue (P = 0.013; Fig. 2A,B). To confirm this finding, we next evaluated ZNF217 protein and mRNA expression in keloid tissue and normal skin tissue by Western blotting and qRT-PCR, respectively. The results showed that ZNF217 was overexpressed in keloid fibroblasts at both the protein and mRNA levels, compared to levels in normal skin tissue (Fig. 2C,D). Altogether, these data suggest that ZNF217 is overexpressed in keloid tissue and fibroblasts.

Bottom Line: Through gain- and loss-of-function studies, we demonstrated that knockdown of lncRNA-ATB decreased autocrine secretion of TGF-β2 and ZNF217 expression but upregulated expression of miR-200c in KFs.Stable downregulation of ZNF217 expression decreased the autocrine secretion of TGF-β2. miR-200c was endogenously associated with lncRNA-ATB, and inhibition of miR-200c overcame the decrease in ZNF217 expression in KFs.Taken together, these findings indicate that lncRNA-ATB governs the autocrine secretion of TGF-β2 in KFs, at least in part, by downregulating the expression level of ZNF217 via miR-200c, suggesting a signaling axis consisting of lncRNA-ATB/miR-200c/ZNF217/TGF-β2.

View Article: PubMed Central - PubMed

Affiliation: Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, Shaanxi, People's Republic of China.

ABSTRACT
Abnormally high activation of transforming growth factor-β (TGF-β) signaling has been demonstrated to be involved in the initiation and progression of keloids. However, the functional role of long non-coding RNA (lncRNA)-activated by TGF-β (lncRNA-ATB) in keloids has not been documented. Here we investigated the role of lncRNA-ATB in the autocrine secretion of TGF-β in keloid fibroblasts (KFs) and explored the underlying molecular mechanism. Using immunohistochemistry and quantitative RT-PCR analysis, we showed that lncRNA-ATB and ZNF217, a transcriptional activator of TGF-β, were overexpressed and miR-200c, which targets ZNF217, was under-expressed in keloid tissue and keloid fibroblasts. Through gain- and loss-of-function studies, we demonstrated that knockdown of lncRNA-ATB decreased autocrine secretion of TGF-β2 and ZNF217 expression but upregulated expression of miR-200c in KFs. Stable downregulation of ZNF217 expression decreased the autocrine secretion of TGF-β2. miR-200c was endogenously associated with lncRNA-ATB, and inhibition of miR-200c overcame the decrease in ZNF217 expression in KFs. Taken together, these findings indicate that lncRNA-ATB governs the autocrine secretion of TGF-β2 in KFs, at least in part, by downregulating the expression level of ZNF217 via miR-200c, suggesting a signaling axis consisting of lncRNA-ATB/miR-200c/ZNF217/TGF-β2. These findings may provide potential biomarkers and targets for novel diagnostic and therapeutic approaches for keloids.

No MeSH data available.


Related in: MedlinePlus