ABSTRACT
This study was designed to evaluate ERK5 expression in lung cancer and malignant melanoma progression and to ascertain the involvement of ERK5 signaling in lung cancer and melanoma. We show that ERK5 expression is abundant in human lung cancer samples, and elevated ERK5 expression in lung cancer was linked to the acquisition of increased metastatic and invasive potential. Importantly, we observed a significant correlation between ERK5 activity and FAK expression and its phosphorylation at the Ser
Subject(s)
Animals , Humans , Mice , A549 Cells , Cell Movement , Epithelial-Mesenchymal Transition/genetics , Focal Adhesion Kinase 1/metabolism , Lung Neoplasms/pathology , MAP Kinase Signaling System , Mitogen-Activated Protein Kinase 7/metabolism , Neoplasm Invasiveness , Neoplasm Metastasis , Neoplasm Proteins/metabolismABSTRACT
Angiotensin II (Ang II) induces the pathological process of vascular structures, including renal glomeruli by hemodynamic and nonhemodynamic direct effects. In kidneys, Ang II plays an important role in the development of proteinuria by the modification of podocyte molecules. We have previously found that Ang II suppressed podocyte AMP-activated protein kinase (AMPK) via Ang II type 1 receptor and MAPK signaling pathway. In the present study, we investigated the roles of AMPK on the changes of p130Cas of podocyte by Ang II. We cultured mouse podocytes and treated them with various concentrations of Ang II and AMPK-modulating agents and analyzed the changes of p130Cas by confocal imaging and western blotting. In immunofluorescence study, Ang II decreased the intensity of p130Cas and changed its localization from peripheral cytoplasm into peri-nuclear areas in a concentrated pattern in podocytes. Ang II also reduced the amount of p130Cas in time and dose-sensitive manners. AMPK activators, metformin and AICAR, restored the suppressed and mal-localized p130Cas significantly, whereas, compound C, an AMPK inhibitor, further aggravated the changes of p130Cas. Losartan, an Ang II type 1 receptor antagonist, recovered the abnormal changes of p130Cas suppressed by Ang II. These results suggest that Ang II induces the relocalization and suppression of podocyte p130Cas by the suppression of AMPK via Ang II type 1 receptor, which would contribute to Ang II-induced podocyte injury.
Subject(s)
Animals , Mice , AMP-Activated Protein Kinases/antagonists & inhibitors , Aminoimidazole Carboxamide/analogs & derivatives , Angiotensin II/pharmacology , Angiotensin II Type 1 Receptor Blockers/pharmacology , Blotting, Western , Cell Line , Cell Nucleus/metabolism , Crk-Associated Substrate Protein/metabolism , Cytoplasm/metabolism , Focal Adhesion Kinase 1/metabolism , Losartan/pharmacology , Metformin/pharmacology , Microscopy, Confocal , Podocytes/cytology , Protein Kinase Inhibitors/pharmacology , Ribonucleotides/pharmacology , Signal Transduction/drug effectsABSTRACT
OBJECTIVE@#To study the expressions of focal adhesion kinase (FAK) and phospho-FAK( p-FAK) during skin incised wound healing and the applicability of time-dependent expressions of FAK and p-FAK.@*METHODS@#The expression of FAK and p-FAK in cutaneous incised wound in mouse were investigated by immunohistochmeistry and Western blotting.@*RESULTS@#FAK and p-FAK expression were detected in polymorphonuclear cells (PMNs) in the wound and adjacent regions 3 hours post-injury. The expressions of FAK and p-FAK were detected in a large number of infiltrating PMNs and some of mononuclear cells (MNCs) from 6 to 24 hours after injury. The MNCs and fibroblastic cells (FBCs) accounted for most part of the FAK and p-FAK positive cells from 3 to 14 days after injury. The numbers of FAK-positive cells increased continuously, reaching a peak at post-injury day 3, and then started to decrease from post-injury day 5 to 14. The changes of p-FAK-positive cells were similar to that of the FAKs, and reached a peak at 12 hours after injury.@*CONCLUSION@#Both FAK and p-FAK displayed a time-dependent expression during skin incised wound healing in mouse, with p-FAK being superior to FAK. Both FAK and p-FAK may potentially be used as new markers for determination of the wound interval.