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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

A schematic model of lncRNA-ATB functions in the pathogenesis of keloids.TGF-β signaling activation regulates a variety of downstream genes including lncRNA-ATB, which sequesters miR-200c away from mRNA of ZNF217, leading to upregulaion of ZNF217 protein and TGF-β2 transcription. Then TGF-β2 binds to TGF-βR and actives TGF-β signaling pathway reversely and forms the LncRNA-ATB/miR-200c/ZNF217/TGF-β2 loop mediating skin fibrosis.
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f7: A schematic model of lncRNA-ATB functions in the pathogenesis of keloids.TGF-β signaling activation regulates a variety of downstream genes including lncRNA-ATB, which sequesters miR-200c away from mRNA of ZNF217, leading to upregulaion of ZNF217 protein and TGF-β2 transcription. Then TGF-β2 binds to TGF-βR and actives TGF-β signaling pathway reversely and forms the LncRNA-ATB/miR-200c/ZNF217/TGF-β2 loop mediating skin fibrosis.

Mentions: In addition, we found that the depletion of lncRNA-ATB, but not lncRNA-508851, decreased ZNF217 expression. Inhibition of miR-200c overcame the decrease in ZNF217 expression (Fig. 6D,E). To ascertain whether this observed effect depends on regulation of the 3′-UTR of ZNF217 by miR-200c, we constructed a luciferase reporter containing the 3′-UTR of ZNF217 (pmirGLO-ZNF217) and transfected it into keloid fibroblasts. The depletion of lncRNA-ATB, but not lncRNA-508851, decreased the luciferase activity of pmirGLO-ZNF217, which was rescued by inhibition of miR-200c (Fig. 6F). All these results suggest that lncRNA-ATB promotes the autocrine secretion of TGF-β2 in keloid fibroblasts via a signaling loop involving lncRNA-ATB/miR-200c/ZNF217/TGF-β2 (Fig. 7).


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)

A schematic model of lncRNA-ATB functions in the pathogenesis of keloids.TGF-β signaling activation regulates a variety of downstream genes including lncRNA-ATB, which sequesters miR-200c away from mRNA of ZNF217, leading to upregulaion of ZNF217 protein and TGF-β2 transcription. Then TGF-β2 binds to TGF-βR and actives TGF-β signaling pathway reversely and forms the LncRNA-ATB/miR-200c/ZNF217/TGF-β2 loop mediating skin fibrosis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835760&req=5

f7: A schematic model of lncRNA-ATB functions in the pathogenesis of keloids.TGF-β signaling activation regulates a variety of downstream genes including lncRNA-ATB, which sequesters miR-200c away from mRNA of ZNF217, leading to upregulaion of ZNF217 protein and TGF-β2 transcription. Then TGF-β2 binds to TGF-βR and actives TGF-β signaling pathway reversely and forms the LncRNA-ATB/miR-200c/ZNF217/TGF-β2 loop mediating skin fibrosis.
Mentions: In addition, we found that the depletion of lncRNA-ATB, but not lncRNA-508851, decreased ZNF217 expression. Inhibition of miR-200c overcame the decrease in ZNF217 expression (Fig. 6D,E). To ascertain whether this observed effect depends on regulation of the 3′-UTR of ZNF217 by miR-200c, we constructed a luciferase reporter containing the 3′-UTR of ZNF217 (pmirGLO-ZNF217) and transfected it into keloid fibroblasts. The depletion of lncRNA-ATB, but not lncRNA-508851, decreased the luciferase activity of pmirGLO-ZNF217, which was rescued by inhibition of miR-200c (Fig. 6F). All these results suggest that lncRNA-ATB promotes the autocrine secretion of TGF-β2 in keloid fibroblasts via a signaling loop involving lncRNA-ATB/miR-200c/ZNF217/TGF-β2 (Fig. 7).

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