Up-regulation of HSP70 by the fibronectin-derived peptide PHSRN in human corneal epithelial cells
Section snippets
Materials and methods
Antibodies and reagents. Mouse monoclonal antibodies to HSP70 and to α-tubulin were obtained from Abcam (Cambridge, MA) and Sigma (St. Louis, MO), respectively. Horseradish peroxidase-conjugated secondary antibodies were obtained from Promega (Madison, WI). SB203580 and PD98059 were obtained from Calbiochem (San Diego, CA), and SP600125 was from Merck (White House Station, NJ). The peptides PHSRN and NRSHP were obtained from Peptide Institute (Osaka, Japan).
Cells. A simian virus 40-transformed
Effects of PHSRN on HSP expression in HCE cells
We first investigated the effects of the PHSRN peptide on the expression of HSP27, HSP47, HSP70, and HSP90 in HCE cells. RT–PCR analysis revealed that the amount of HSP70 mRNA in HCE cells exposed to PHSRN (1 μg/ml) for 12 h was markedly increased compared with that in cells exposed to the control peptide NRSHP or incubated alone (Fig. 1). In contrast, PHSRN had no effect on the amounts of HSP27, HSP47, or HSP90 mRNAs.
The effect of PHSRN on the abundance of HSP70 mRNA was concentration dependent,
Discussion
We have shown that the PHSRN peptide up-regulated expression of HSP70, but not that of other HSPs (HSP27, HSP47, HSP90), in HCE cells. This effect of PHSRN was apparent at both the protein and mRNA levels and appeared to be mediated by the p38 MAPK signaling pathway, but not by the ERK or JNK signaling pathways. Our results thus suggest that up-regulation of HSP70 expression by PHSRN, corresponding to the second cell-binding domain of fibronectin, may contribute to the stimulation of corneal
Acknowledgments
We thank Shizuka Murata, Yukari Mizuno, and Yasumiko Akamatsu for technical assistance.
References (26)
- et al.
Role of lumican in the corneal epithelium during wound healing
J. Biol. Chem.
(2000) - et al.
Expression of HSP70 in healing wounds of diabetic and nondiabetic mice
J. Surg. Res.
(1999) - et al.
Differential expression of heat shock protein 70 in well healing and chronic human wound tissue
Biochem. Biophys. Res. Commun.
(1995) The role of heat-shock proteins as molecular chaperones
Curr. Opin. Cell Biol.
(1991)- et al.
Involvement of LOX-1 in dendritic cell-mediated antigen cross-presentation
Immunity
(2002) - et al.
Human HSP 27 is phosphorylated at serines 78 and 82 by heat shock and mitogen-activated kinases that recognize the same amino acid motif as S6 kinase II
J. Biol. Chem.
(1992) - et al.
Differential activation of p38 mitogen-activated protein kinase and extracellular signal-regulated protein kinases confers cadmium-induced HSP 70 expression in 9L rat brain tumor cells
J. Biol. Chem.
(1998) - et al.
Fibronectin in healing rabbit corneal wounds
Lab. Invest.
(1981) - et al.
Fibronectin in corneal wound healing: appearance in cultured rabbit cornea
Jpn. J. Ophthalmol.
(1982) - et al.
Production of fibronectin and tenascin isoforms and their role in the adhesion of human immortalized corneal epithelial cells
Invest. Ophthalmol. Vis. Sci.
(2003)
Enhancement of vinculin synthesis by migrating stratified squamous epithelium
J. Cell Biol.
The human cornea proteome: bioinformatic analysis indicate import of plasma proteins into the cornea
J. Mol. Vis.
Stimulation of corneal epithelial migration by a synthetic peptide (PHSRN) corresponding to the second cell-binding site of fibronectin
Invest. Ophthalmol. Vis. Sci.
Cited by (17)
Wound healing in the eye: Therapeutic prospects
2018, Advanced Drug Delivery ReviewsCitation Excerpt :The amino acid sequence Pro-His-Ser-Arg-Asn (PHSRN) is thought to be a secondary binding site of fibronectin [146] as it mirrors many of the effects that fibronectin has on corneal epithelial cells [147]. This fibronectin-derived peptide sequence has been shown to upregulate the expression of the tight-junction protein Zonula occludens-1 (ZO-1) as well as heat shock protein 70 by activating protein kinase signaling pathways in the corneal epithelium [148,149]. PHSRN treatment has been reported to stimulate corneal epithelial migration in vitro [147].
Aspergillus flavus induced alterations in tear protein profile reveal pathogen-induced host response to fungal infection
2017, Journal of ProteomicsCitation Excerpt :Up-regulation of plasminogen, whose active form can clear fibrin clot suggests a fine balance in clot formation and clearance. Up-regulation of three heat shock proteins, HSP beta 1 (Hsp27), Hsp70 (1A variant), and Hsp90 alpha in tear from keratitis patients imply that these proteins may have a role in wound healing in addition to chaperone function as has been shown in previous studies [43–46]. The presence of lumican, vimentin (a pro-fibrotic protein), and MMP-9 in the patient tear indicate the presence of yet another set of factors required for wound healing process.
Peptide therapies for ocular surface disturbances based on fibronectin-integrin interactions
2015, Progress in Retinal and Eye ResearchCitation Excerpt :PHSRN also increased the levels of HSP70 (heat shock protein of 70 kDa) mRNA and protein in these cells in a concentration- and time-dependent manner, whereas NRSHP did not. The PHSRN-induced up-regulation of HSP70 was blocked by SB203580, an inhibitor of p38 mitogen-activated protein kinase (MAPK), but it was not affected by PD98059 or SP600125, inhibitors of signaling by the MAPKs ERK and c-Jun NH2-terminal kinase (JNK), respectively (Ko et al., 2008c). These results suggested that the induction of HSP70 expression might contribute to the promotion of corneal epithelial migration by PHSRN and hence to corneal epithelial wound healing.
P38 MAPK inhibition alleviates experimental acute pancreatitis in mice
2015, Hepatobiliary and Pancreatic Diseases International