Limits...
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

Stable knockdown of lncRNA-ATB inhibits autocrine secretion of TGF-β2 in keloid fibroblast.(A,B) qRT-PCR analysis of lncRNA-ATB levels in keloid (K) and normal skin (NS) tissues (57 paired samples), as well as in 17 pairs of keloid fibroblasts (KF) and normal fibroblasts (NF); (C) qRT-PCR determination of relative lncRNA-ATB expression levels in KF at 48 h and 72 h after knockdown of lncRNA-ATB with lncRNA-ATB–specific shRNAs (shRNA-ATB-1 and shRNA-ATB-2) compared with a negative control siRNA (shRNA-control); (D) ELISA of relative TGF-β2 expression levels in KF at 48 h and 72 h after knockdown of lncRNA-ATB.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4835760&req=5

f1: Stable knockdown of lncRNA-ATB inhibits autocrine secretion of TGF-β2 in keloid fibroblast.(A,B) qRT-PCR analysis of lncRNA-ATB levels in keloid (K) and normal skin (NS) tissues (57 paired samples), as well as in 17 pairs of keloid fibroblasts (KF) and normal fibroblasts (NF); (C) qRT-PCR determination of relative lncRNA-ATB expression levels in KF at 48 h and 72 h after knockdown of lncRNA-ATB with lncRNA-ATB–specific shRNAs (shRNA-ATB-1 and shRNA-ATB-2) compared with a negative control siRNA (shRNA-control); (D) ELISA of relative TGF-β2 expression levels in KF at 48 h and 72 h after knockdown of lncRNA-ATB.

Mentions: Because lncRNA-ATB has been demonstrated to play an important role in TGF-β signaling in hepatocellular carcinoma7, we investigated whether lncRNA-ATB also acts similarly in keloids and keloid fibroblasts, which play a critical role in the progression of keloids. As anticipated, qRT-PCR analysis showed that lncRNA-ATB expression was significantly increased in both keloid tissue and keloid fibroblasts compared to normal skin tissue and normal fibroblasts, respectively (Fig. 1A,B).


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)

Stable knockdown of lncRNA-ATB inhibits autocrine secretion of TGF-β2 in keloid fibroblast.(A,B) qRT-PCR analysis of lncRNA-ATB levels in keloid (K) and normal skin (NS) tissues (57 paired samples), as well as in 17 pairs of keloid fibroblasts (KF) and normal fibroblasts (NF); (C) qRT-PCR determination of relative lncRNA-ATB expression levels in KF at 48 h and 72 h after knockdown of lncRNA-ATB with lncRNA-ATB–specific shRNAs (shRNA-ATB-1 and shRNA-ATB-2) compared with a negative control siRNA (shRNA-control); (D) ELISA of relative TGF-β2 expression levels in KF at 48 h and 72 h after knockdown of lncRNA-ATB.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: Stable knockdown of lncRNA-ATB inhibits autocrine secretion of TGF-β2 in keloid fibroblast.(A,B) qRT-PCR analysis of lncRNA-ATB levels in keloid (K) and normal skin (NS) tissues (57 paired samples), as well as in 17 pairs of keloid fibroblasts (KF) and normal fibroblasts (NF); (C) qRT-PCR determination of relative lncRNA-ATB expression levels in KF at 48 h and 72 h after knockdown of lncRNA-ATB with lncRNA-ATB–specific shRNAs (shRNA-ATB-1 and shRNA-ATB-2) compared with a negative control siRNA (shRNA-control); (D) ELISA of relative TGF-β2 expression levels in KF at 48 h and 72 h after knockdown of lncRNA-ATB.
Mentions: Because lncRNA-ATB has been demonstrated to play an important role in TGF-β signaling in hepatocellular carcinoma7, we investigated whether lncRNA-ATB also acts similarly in keloids and keloid fibroblasts, which play a critical role in the progression of keloids. As anticipated, qRT-PCR analysis showed that lncRNA-ATB expression was significantly increased in both keloid tissue and keloid fibroblasts compared to normal skin tissue and normal fibroblasts, respectively (Fig. 1A,B).

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