title: Expression and intracellular localization of Smad proteins in human endometrial cancer.
abstract: The aim of our study was to examine expression of Smad proteins i.e., Smad2, Smad3 and Smad4 both as mRNA and protein as well as their intracellular localization in normal (n=13) and neoplastic (n=42) endometrial tissue specimens using RT-PCR and immunological techniques i.e., Western blot and ELISA. Two uncommon female genital tract tumours, rhabdomyosarcoma of uterine of the cervix and uterine carcinosarcoma were also included. No statistically significant differences were found in the mRNA level of the examined Smad proteins between normal and tumour tissue specimens. Smad2 and Smad3 mRNAs were detected both in uterine carcinosarcoma and rhabdomyosarcoma of the uterine cervix. However, significantly lower Smad2 and Smad4 mRNA level was noted when the depth of myometrial invasion was considered (p<0.05). In endometrial cancer as compared to normal endometrium significantly higher levels of Smad2 and Smad3 proteins, both in cytoplasmic (p=0.002; p=0.0001) and nuclear (p=0.016; p=0.0004) fractions were observed. Both in uterine carcinosarcoma and rhabdomyosarcoma of the uterine cervix Smad2, Smad3 and Smad4 proteins were not detected. Moreover, significantly elevated Smad4 protein level in cytoplasmic fraction was stated when tumour grade and depth of myometrial invasion was undertaken (p<0.05). When intracellular distribution of Smads was considered differences between cytoplasmic and nuclear localization in normal and carcinomatous endometrium was stated. In endometrial cancer decreased number of cases with Smad3 and increased number of cases with Smad4 located in nuclear fraction was found. In conclusion, the disturbances in Smad protein expression and/or differences in their intracellular distribution suggest, that TGF-beta signaling pathway via Smads may be deregulated in endometrial carcinomas.
authors.affiliation: Department of Cytobiochemistry, University of Lodz, 90-237 Lodz, Poland. kalaiste@wp.pl
authors.forename: Dagmara
authors.initials: D
authors.lastname: Piestrzeniewicz-Ulanska
authors.forename: Magdalena
authors.initials: M
authors.lastname: Brys
authors.forename: Andrzej
authors.initials: A
authors.lastname: Semczuk
authors.forename: Jerzy A
authors.initials: JA
authors.lastname: Jakowicki
authors.forename: Wanda M
authors.initials: WM
authors.lastname: Krajewska
mesh_terms: D015153:Blotting, Western; D002296:Carcinosarcoma / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D016889:Endometrial Neoplasms / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012208:Rhabdomyosarcoma / Q000378:metabolism; D051785:Smad Proteins; D051901:Smad4 Protein; D013347:Subcellular Fractions; D015534:Trans-Activators / Q000096:biosynthesis*; D014594:Uterine Neoplasms / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; C497796:SMAD4 protein, human; D051785:Smad Proteins; D051901:Smad4 Protein; D015534:Trans-Activators