title: Expression, localization, and phosphorylation of Akt1 in benign and malignant thyroid lesions.
abstract: The serine/threonine protein kinase Akt is a key molecule in the phosphatidyl inositol 3-kinase pathway that is often overactivated in human cancers. Three Akt isoforms (Akt1, Akt2, Akt3) have been identified in human cells and they show different distribution and have non-redundant functions. The aim of this study was to determine whether the expression, phosphorylation, and localization of Akt1 isoform in human thyroid malignant lesions are different from those in benign lesions. Nuclear and cytoplasmic fractions were isolated from tissue samples and Western blot method was used to detect Akt1 presence in both cellular fractions. Akt1 expression was also assessed by ELISA method. To estimate Akt1 phosphorylation, kinase was immunoprecipitated from cell lysates and tested with anti-phospho-Akt antibodies. The Akt1 expression in majority of thyroid cancer samples was significantly higher than in benign lesions (p < 0.05). Akt1 both in differentiated cancers (follicular and papillary) and benign lesions was localized mainly in cytoplasmic fraction. In two of three anaplastic cancer samples Akt1 was predominantly localized in nucleus. The ratio of phosphorylated Akt1 to total Akt1 was lower in cancers than in non-neoplastic lesions and adenomas. Thus, although Akt1 seems to be overexpressed in thyroid neoplasms, its high phosphorylation is not characteristic for thyroid cancers.
authors: Department of Cytobiochemistry, University of Lodz, Pomorska 141/143, 90-236, Lodz, Poland. krzeslaka@interia.pl
authors: Anna
authors: A
authors: Krześlak
authors: Lech
authors: L
authors: Pomorski
authors: Anna
authors: A
authors: Lipińska
mesh_terms: D015153:Blotting, Western; D018265:Carcinoma, Papillary, Follicular / Q000201:enzymology*; D002458:Cell Fractionation; D003593:Cytoplasm / Q000201:enzymology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D008297:Male; D009687:Nuclear Proteins / Q000378:metabolism; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000096:biosynthesis / Q000378:metabolism*; D015398:Signal Transduction; D018709:Statistics, Nonparametric; D013964:Thyroid Neoplasms / Q000201:enzymology*
chemical_list: D009687:Nuclear Proteins; C494918:AKT1 protein, human; D051057:Proto-Oncogene Proteins c-akt