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Upregulated periostin promotes angiogenesis in keloids through activation of the ERK 1/2 and focal adhesion kinase pathways, as well as the upregulated expression of VEGF and angiopoietin‑1.

Zhang Z, Nie F, Chen X, Qin Z, Kang C, Chen B, Ma J, Pan B, Ma Y - Mol Med Rep (2014)

Bottom Line: Conditioned medium from keloid fibroblasts (KFs) promoted the migration and tube formation of human umbilical vein endothelial cells (HUVECs) compared with normal fibroblasts and this effect may have been abrogated by the short hairpin RNA knockdown of periostin.Treatment with recombinant human periostin promoted the migration and tube formation of HUVECs by activating the extracellular signal‑regulated kinase 1/2 and focal adhesion kinase signaling pathway.In addition, periostin increased the secretion of vascular endothelial growth factor and angiopoietin‑1 in the KFs.

View Article: PubMed Central - PubMed

Affiliation: Department of Plastic Surgery, Peking University Third Hospital, Beijing 100191, P.R. China.

ABSTRACT
Periostin, a secreted extracellular matrix protein, is highly expressed in wound healing and in various types of human cancer and is involved in angiogenesis. Keloids, considered dermal benign tumors, are granulomatous lesions characterized by capillary proliferation. However, the underlying regulatory mechanism of angiogenesis in keloids remains to be elucidated. The present study aimed to examine the effect of periostin on angiogenesis in keloids. The expression of periostin was upregulated and the vessel density was higher in human keloids compared with normal tissue, observed following staining with CD31 and CD105. Periostin demonstrated a markedly positive correlation with blood vessel density, which was assessed using CD31 staining (r=0.711; P<0.01) and a weak correlation was observed using CD105 staining (r=0.251; P<0.01). Conditioned medium from keloid fibroblasts (KFs) promoted the migration and tube formation of human umbilical vein endothelial cells (HUVECs) compared with normal fibroblasts and this effect may have been abrogated by the short hairpin RNA knockdown of periostin. Treatment with recombinant human periostin promoted the migration and tube formation of HUVECs by activating the extracellular signal‑regulated kinase 1/2 and focal adhesion kinase signaling pathway. In addition, periostin increased the secretion of vascular endothelial growth factor and angiopoietin‑1 in the KFs. In conclusion, these data suggested that upregulation in the level of periostin may promote angiogenesis directly and indirectly in keloids and may be a key factor in keloid development. Periostin may, therefore, be a promising therapeutic target in the treatment of keloids and other angioproliferative diseases.

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Lentiviral-mediated stable genetic knockdown in KFs. KFs were transfected with lentivirus harboring shRNA periostin or non-silencing shRNA. (A) Fluorescence microscopy of sh-pn under (a) microscopy and (b) fluorescence microscopy; non-silencing group under (c) microscopy and (d) fluorescence microscopy. (B) mRNA analyses were performed to ensure the efficiency of transfection prior to each experiment (n=6). *P<0.05, compared with the controls. n.s,, not significant; KF, keloid fibroblasts; shRNA, short hairpin RNA; sh-pm, shRNA-periostin; NC, non-silencing.
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f2-mmr-11-02-0857: Lentiviral-mediated stable genetic knockdown in KFs. KFs were transfected with lentivirus harboring shRNA periostin or non-silencing shRNA. (A) Fluorescence microscopy of sh-pn under (a) microscopy and (b) fluorescence microscopy; non-silencing group under (c) microscopy and (d) fluorescence microscopy. (B) mRNA analyses were performed to ensure the efficiency of transfection prior to each experiment (n=6). *P<0.05, compared with the controls. n.s,, not significant; KF, keloid fibroblasts; shRNA, short hairpin RNA; sh-pm, shRNA-periostin; NC, non-silencing.

Mentions: To determine whether the periostin secreted by fibroblasts affected angiogenesis, the present study examined the migration and tube formation of HUVECs using conditioned medium from the KFs and NFs. Firstly, the KFs were transfected with either shRNA against periostin or control shRNA. Fluorescence microscopy and reverse transcription quantitative polymerase chain reaction (RT-qPCR) analyses were performed to ensure the efficiency of transfection prior to each experiment (Fig. 2). Use of the KF-conditioned medium significantly increased the number of migrating cells compared with the NF-conditioned medium (Fig. 3A). In addition, KF-conditioned medium promoted tube formation when compared with the NF-conditioned medium (Fig. 3B). Notably, the conditioned medium from the KFs with shRNA knock down of periostin significantly inhibited migration and tube formation compared with the controls. To exclude the effect of other factors in the conditioned medium, the effect of rhPN treatment at different concentrations on HUVEC migration and tube formation was examined. Contrary to the effects of conditioned medium from the KFs with shRNA periostin-knockdown, treatment with rhPN dose-dependently promoted migration (Fig. 4A) and tube formation (Fig. 4B).


Upregulated periostin promotes angiogenesis in keloids through activation of the ERK 1/2 and focal adhesion kinase pathways, as well as the upregulated expression of VEGF and angiopoietin‑1.

Zhang Z, Nie F, Chen X, Qin Z, Kang C, Chen B, Ma J, Pan B, Ma Y - Mol Med Rep (2014)

Lentiviral-mediated stable genetic knockdown in KFs. KFs were transfected with lentivirus harboring shRNA periostin or non-silencing shRNA. (A) Fluorescence microscopy of sh-pn under (a) microscopy and (b) fluorescence microscopy; non-silencing group under (c) microscopy and (d) fluorescence microscopy. (B) mRNA analyses were performed to ensure the efficiency of transfection prior to each experiment (n=6). *P<0.05, compared with the controls. n.s,, not significant; KF, keloid fibroblasts; shRNA, short hairpin RNA; sh-pm, shRNA-periostin; NC, non-silencing.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4262479&req=5

f2-mmr-11-02-0857: Lentiviral-mediated stable genetic knockdown in KFs. KFs were transfected with lentivirus harboring shRNA periostin or non-silencing shRNA. (A) Fluorescence microscopy of sh-pn under (a) microscopy and (b) fluorescence microscopy; non-silencing group under (c) microscopy and (d) fluorescence microscopy. (B) mRNA analyses were performed to ensure the efficiency of transfection prior to each experiment (n=6). *P<0.05, compared with the controls. n.s,, not significant; KF, keloid fibroblasts; shRNA, short hairpin RNA; sh-pm, shRNA-periostin; NC, non-silencing.
Mentions: To determine whether the periostin secreted by fibroblasts affected angiogenesis, the present study examined the migration and tube formation of HUVECs using conditioned medium from the KFs and NFs. Firstly, the KFs were transfected with either shRNA against periostin or control shRNA. Fluorescence microscopy and reverse transcription quantitative polymerase chain reaction (RT-qPCR) analyses were performed to ensure the efficiency of transfection prior to each experiment (Fig. 2). Use of the KF-conditioned medium significantly increased the number of migrating cells compared with the NF-conditioned medium (Fig. 3A). In addition, KF-conditioned medium promoted tube formation when compared with the NF-conditioned medium (Fig. 3B). Notably, the conditioned medium from the KFs with shRNA knock down of periostin significantly inhibited migration and tube formation compared with the controls. To exclude the effect of other factors in the conditioned medium, the effect of rhPN treatment at different concentrations on HUVEC migration and tube formation was examined. Contrary to the effects of conditioned medium from the KFs with shRNA periostin-knockdown, treatment with rhPN dose-dependently promoted migration (Fig. 4A) and tube formation (Fig. 4B).

Bottom Line: Conditioned medium from keloid fibroblasts (KFs) promoted the migration and tube formation of human umbilical vein endothelial cells (HUVECs) compared with normal fibroblasts and this effect may have been abrogated by the short hairpin RNA knockdown of periostin.Treatment with recombinant human periostin promoted the migration and tube formation of HUVECs by activating the extracellular signal‑regulated kinase 1/2 and focal adhesion kinase signaling pathway.In addition, periostin increased the secretion of vascular endothelial growth factor and angiopoietin‑1 in the KFs.

View Article: PubMed Central - PubMed

Affiliation: Department of Plastic Surgery, Peking University Third Hospital, Beijing 100191, P.R. China.

ABSTRACT
Periostin, a secreted extracellular matrix protein, is highly expressed in wound healing and in various types of human cancer and is involved in angiogenesis. Keloids, considered dermal benign tumors, are granulomatous lesions characterized by capillary proliferation. However, the underlying regulatory mechanism of angiogenesis in keloids remains to be elucidated. The present study aimed to examine the effect of periostin on angiogenesis in keloids. The expression of periostin was upregulated and the vessel density was higher in human keloids compared with normal tissue, observed following staining with CD31 and CD105. Periostin demonstrated a markedly positive correlation with blood vessel density, which was assessed using CD31 staining (r=0.711; P<0.01) and a weak correlation was observed using CD105 staining (r=0.251; P<0.01). Conditioned medium from keloid fibroblasts (KFs) promoted the migration and tube formation of human umbilical vein endothelial cells (HUVECs) compared with normal fibroblasts and this effect may have been abrogated by the short hairpin RNA knockdown of periostin. Treatment with recombinant human periostin promoted the migration and tube formation of HUVECs by activating the extracellular signal‑regulated kinase 1/2 and focal adhesion kinase signaling pathway. In addition, periostin increased the secretion of vascular endothelial growth factor and angiopoietin‑1 in the KFs. In conclusion, these data suggested that upregulation in the level of periostin may promote angiogenesis directly and indirectly in keloids and may be a key factor in keloid development. Periostin may, therefore, be a promising therapeutic target in the treatment of keloids and other angioproliferative diseases.

Show MeSH
Related in: MedlinePlus