BACKGROUND: In human epidermal keratinocytes the expression of hsp27 is closely related to differentiation in vitro and in situ. OBJECTIVE: We aimed to gain further insight into the role of hsp27 in epidermal differentiation by specific inhibition through siRNA and inhibition of p38-MAPK, the key enzyme of hsp27 phosphorylation. METHODS: Normal human keratinocytes (KC) and organotypic skin cultures (SE-skin equivalents) were used. Expression and phosphorylation of hsp27 was inhibited in these models by siRNA and SB203580, a specific inhibitor of p38-MAPK, respectively. Modification of morphology and expression of hsp27 and other differentiation associated proteins was investigated by immunofluorescence, western blot, and RT-PCR. RESULTS: Inhibition of p38-MAPK resulted in a downregulation of hsp27 in KC and SE. Additionally, in the presence of SB203580 Ca(2+) induced expression of pro-filaggrin and loricrin was inhibited at the protein level and expression of filaggrin, keratin 10, and transglutaminase 1 at the mRNA level. Addition of SB203580 to SE, as well as hsp27 knockdown in this model resulted in identical patterns of irregular differentiation, disturbance of epidermal layers, and delayed expression of K10. CONCLUSION: These results provide evidence that the expression of hsp27 and its phosphorylation by p38-MAPK are required for keratinocyte differentiation and for the formation of a regularly stratified epidermis. Copyright Â
BACKGROUND: In human epidermal keratinocytes the expression of hsp27 is closely related to differentiation in vitro and in situ. OBJECTIVE: We aimed to gain further insight into the role of hsp27 in epidermal differentiation by specific inhibition through siRNA and inhibition of p38-MAPK, the key enzyme of hsp27 phosphorylation. METHODS: Normal human keratinocytes (KC) and organotypic skin cultures (SE-skin equivalents) were used. Expression and phosphorylation of hsp27 was inhibited in these models by siRNA and SB203580, a specific inhibitor of p38-MAPK, respectively. Modification of morphology and expression of hsp27 and other differentiation associated proteins was investigated by immunofluorescence, western blot, and RT-PCR. RESULTS: Inhibition of p38-MAPK resulted in a downregulation of hsp27 in KC and SE. Additionally, in the presence of SB203580Ca(2+) induced expression of pro-filaggrin and loricrin was inhibited at the protein level and expression of filaggrin, keratin 10, and transglutaminase 1 at the mRNA level. Addition of SB203580 to SE, as well as hsp27 knockdown in this model resulted in identical patterns of irregular differentiation, disturbance of epidermal layers, and delayed expression of K10. CONCLUSION: These results provide evidence that the expression of hsp27 and its phosphorylation by p38-MAPK are required for keratinocyte differentiation and for the formation of a regularly stratified epidermis. Copyright Â
Authors: Rong Wang; Joan D Ferraris; Yuichiro Izumi; Natalia Dmitrieva; Kevin Ramkissoon; Guanghui Wang; Marjan Gucek; Maurice B Burg Journal: Am J Physiol Cell Physiol Date: 2014-06-25 Impact factor: 4.249
Authors: Leonardo Almeida-Souza; Bob Asselbergh; Vicky De Winter; Sofie Goethals; Vincent Timmerman; Sophie Janssens Journal: PLoS One Date: 2013-06-24 Impact factor: 3.240
Authors: Katrina M Waters; Brian S Cummings; Harish Shankaran; Natalie E Scholpa; Thomas J Weber Journal: Chem Res Toxicol Date: 2014-08-12 Impact factor: 3.739