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Role for heat shock protein 90α in the proliferation and migration of HaCaT cells and in the deep second-degree burn wound healing in mice.

Zhang Y, Bai X, Wang Y, Li N, Li X, Han F, Su L, Hu D - PLoS ONE (2014)

Bottom Line: The hypoxic wound microenvironment promotes cell migration through a hypoxia--heat shock protein 90 alpha (Hsp90α)--low density lipoprotein receptor-related protein-1 (LRP-1) autocrine loop.Experiments performed with a human keratinocyte cell line--HaCaT also confirmed above results. 17-dimethylaminoethylamino-17demethoxygeldanamycin hydrochloride (17-DMAG), an Hsp90α inhibitor, was used to further evaluate the function of Hsp90α in wound healing.In conclusion, these results provided a rationale for the therapeutic effect of Hsp90α on the burn wound management.

View Article: PubMed Central - PubMed

Affiliation: Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shannxi, China.

ABSTRACT
Inflammation, proliferation, and tissue remodeling are essential steps for wound healing. The hypoxic wound microenvironment promotes cell migration through a hypoxia--heat shock protein 90 alpha (Hsp90α)--low density lipoprotein receptor-related protein-1 (LRP-1) autocrine loop. To elucidate the role of this autocrine loop on burn wound healing, we investigated the expression profile of Hsp90α at the edge of burn wounds and found a transient increase in both mRNA and protein levels. Experiments performed with a human keratinocyte cell line--HaCaT also confirmed above results. 17-dimethylaminoethylamino-17demethoxygeldanamycin hydrochloride (17-DMAG), an Hsp90α inhibitor, was used to further evaluate the function of Hsp90α in wound healing. Consistently, topical application of Hsp90α in the early stage of deep second-degree burn wounds led to reduced inflammation and increased tissue granulation, with a concomitant reduction in the size of the wound at each time point tested (p<0.05). Consequently, epidermal cells at the wound margin progressed more rapidly causing an expedited healing process. In conclusion, these results provided a rationale for the therapeutic effect of Hsp90α on the burn wound management.

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Flow cytometry assay showing the effects of Hsp90α on cell cycle progression and apoptosis following heat shock.Heat-shocked cells treated with (A) saline, (B) Hsp90α, and (C) 17-DMAG for 24 h were assessed by flow cytometry. Hsp90α increased the number of cells in G1 phase, while 17-DMAG induced cell cycle arrest at G0–G1 phases. These results were represented histogramatically in (D). FITC- Annexin V/propidium iodide (PI) was used to measure the apoptosis rate of HaCaT cells following three treatments: (E) saline, (F) Hsp90α, and (G)17-DMAG. The percentage of apoptotic cell population was decreased in Hsp90α group and increased in 17-DMAG group (H) (p<0.05).
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pone-0103723-g005: Flow cytometry assay showing the effects of Hsp90α on cell cycle progression and apoptosis following heat shock.Heat-shocked cells treated with (A) saline, (B) Hsp90α, and (C) 17-DMAG for 24 h were assessed by flow cytometry. Hsp90α increased the number of cells in G1 phase, while 17-DMAG induced cell cycle arrest at G0–G1 phases. These results were represented histogramatically in (D). FITC- Annexin V/propidium iodide (PI) was used to measure the apoptosis rate of HaCaT cells following three treatments: (E) saline, (F) Hsp90α, and (G)17-DMAG. The percentage of apoptotic cell population was decreased in Hsp90α group and increased in 17-DMAG group (H) (p<0.05).

Mentions: Hsp90α is known to function as a chaperone for a variety of proteins that regulate cell cycle progression and apoptosis [27], [28]. To determine the effect of Hsp90α on the viability of heat-shocked cells, we evaluated the cell cycle distribution and the level of HaCaT cell apoptosis by flow cytometry. As shown in Fig. 5A–D, heat-shocked cells treated with Hsp90α exhibited an obvious decrease in the cell number of G1 phrase, while 17-DMAG showed an inductive effect on G1 cell cycle arrest. Furthermore, as shown in Fig. 5E–H, the apoptosis rate of HaCaT cells was significantly decreased after Hsp90α treatment, while 17-DMAG treatment showed the opposite effect. These results further illustrated the role of Hsp90α on the viability and apoptosis of heat-shocked cells.


Role for heat shock protein 90α in the proliferation and migration of HaCaT cells and in the deep second-degree burn wound healing in mice.

Zhang Y, Bai X, Wang Y, Li N, Li X, Han F, Su L, Hu D - PLoS ONE (2014)

Flow cytometry assay showing the effects of Hsp90α on cell cycle progression and apoptosis following heat shock.Heat-shocked cells treated with (A) saline, (B) Hsp90α, and (C) 17-DMAG for 24 h were assessed by flow cytometry. Hsp90α increased the number of cells in G1 phase, while 17-DMAG induced cell cycle arrest at G0–G1 phases. These results were represented histogramatically in (D). FITC- Annexin V/propidium iodide (PI) was used to measure the apoptosis rate of HaCaT cells following three treatments: (E) saline, (F) Hsp90α, and (G)17-DMAG. The percentage of apoptotic cell population was decreased in Hsp90α group and increased in 17-DMAG group (H) (p<0.05).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4128658&req=5

pone-0103723-g005: Flow cytometry assay showing the effects of Hsp90α on cell cycle progression and apoptosis following heat shock.Heat-shocked cells treated with (A) saline, (B) Hsp90α, and (C) 17-DMAG for 24 h were assessed by flow cytometry. Hsp90α increased the number of cells in G1 phase, while 17-DMAG induced cell cycle arrest at G0–G1 phases. These results were represented histogramatically in (D). FITC- Annexin V/propidium iodide (PI) was used to measure the apoptosis rate of HaCaT cells following three treatments: (E) saline, (F) Hsp90α, and (G)17-DMAG. The percentage of apoptotic cell population was decreased in Hsp90α group and increased in 17-DMAG group (H) (p<0.05).
Mentions: Hsp90α is known to function as a chaperone for a variety of proteins that regulate cell cycle progression and apoptosis [27], [28]. To determine the effect of Hsp90α on the viability of heat-shocked cells, we evaluated the cell cycle distribution and the level of HaCaT cell apoptosis by flow cytometry. As shown in Fig. 5A–D, heat-shocked cells treated with Hsp90α exhibited an obvious decrease in the cell number of G1 phrase, while 17-DMAG showed an inductive effect on G1 cell cycle arrest. Furthermore, as shown in Fig. 5E–H, the apoptosis rate of HaCaT cells was significantly decreased after Hsp90α treatment, while 17-DMAG treatment showed the opposite effect. These results further illustrated the role of Hsp90α on the viability and apoptosis of heat-shocked cells.

Bottom Line: The hypoxic wound microenvironment promotes cell migration through a hypoxia--heat shock protein 90 alpha (Hsp90α)--low density lipoprotein receptor-related protein-1 (LRP-1) autocrine loop.Experiments performed with a human keratinocyte cell line--HaCaT also confirmed above results. 17-dimethylaminoethylamino-17demethoxygeldanamycin hydrochloride (17-DMAG), an Hsp90α inhibitor, was used to further evaluate the function of Hsp90α in wound healing.In conclusion, these results provided a rationale for the therapeutic effect of Hsp90α on the burn wound management.

View Article: PubMed Central - PubMed

Affiliation: Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shannxi, China.

ABSTRACT
Inflammation, proliferation, and tissue remodeling are essential steps for wound healing. The hypoxic wound microenvironment promotes cell migration through a hypoxia--heat shock protein 90 alpha (Hsp90α)--low density lipoprotein receptor-related protein-1 (LRP-1) autocrine loop. To elucidate the role of this autocrine loop on burn wound healing, we investigated the expression profile of Hsp90α at the edge of burn wounds and found a transient increase in both mRNA and protein levels. Experiments performed with a human keratinocyte cell line--HaCaT also confirmed above results. 17-dimethylaminoethylamino-17demethoxygeldanamycin hydrochloride (17-DMAG), an Hsp90α inhibitor, was used to further evaluate the function of Hsp90α in wound healing. Consistently, topical application of Hsp90α in the early stage of deep second-degree burn wounds led to reduced inflammation and increased tissue granulation, with a concomitant reduction in the size of the wound at each time point tested (p<0.05). Consequently, epidermal cells at the wound margin progressed more rapidly causing an expedited healing process. In conclusion, these results provided a rationale for the therapeutic effect of Hsp90α on the burn wound management.

Show MeSH
Related in: MedlinePlus