(55-57) Immunohistochemical studies using HBME-1, galectin-3, CITED1, cytokeratin 19 (CK19), and cyclin D1 in these borderline follicular lesions have shown heterogeneous and intermediate staining patterns compared with PTC, suggesting a potential relationship to carcinoma and a role as precursor lesions.
Ma et al (88) showed increased expression of PTC-associated markers, such as neutrophil gelatinase-associated lipocalin, CK19, and claudin 1, in foci of atypical follicular epithelium compared with peritumoral benign thyroid tissue, whereas Prasad et al (89) identified the expression of HBME-1, galectin-3, CK19, CITED1, and fibronectin in PTC-like nuclear alterations of CLT.
Compared to SAT, the levels of PGC-1[alpha], UCP-1, CITED1, and TFAM, which play major roles in mitochondrial activity and energy expenditure in brown adipocytes, increased significantly (Figures 3(a), 3(b), and 3(c)).
The levels of proteins involved in energy expenditure (PGC-1[alpha], UCP-1, CITED1, and TFAM) increased in adipocytes from PAT compared to those from SAT.
Membranes were then incubated with antibodies targeting proteins involved in thermogenesis: rabbit anti-PGC-1[alpha] (1:1000, Abcam ab54481), rabbit anti-UCP-1 (1:1000, Abcam ab155117), rabbit anti-IL-4R (1:1000, Abcam ab131058), rabbit anti-STAT6 phosphorylated (phospho Y641) (1: 1000, Abcam ab54461), and mouse anti-cAMP response element-binding protein/p300-interacting transactivator 1 (CITED1; Abcam ab87978).
UCP-1 mRNA was shown to be induced in the presence of IL-4 (Figure 3(a)), and, as shown in Figures 3(b) and 3(c), protein expression levels of PGC-1a and CITED1 were increased by 2.3- and nearly 2-fold, respectively.
In embryonic kidney, the transcription factor sine oculis homeobox homolog 2 (SIX-2) and aspartic acid rich C terminal domain 1 (CITED1) that are essential for self-renewal are considered as markers of MMSCs [14, 22].
Western blotting was performed to examine CITED1, SIX-2, E-cadherin, and the related proteins in the Wnt/[beta]-Catenin signalling pathway (Wnt4, [beta]-catenin, and LEF1).
Some of these useful antibodies in the diagnosis of FVPTC include cytokeratin 19 (CK19), Hector Battifora mesothelial 1 (HBME1), CREB-binding protein/p300-interacting transactivator with Asp/Glu-rich C-terminal domain 1 (CITED1), galactoside-binding lectin soluble 3 (Galectin-3), CD57, retinoblastoma (Rb) protein, and p27, among others (31,43-46) (Table).
Abbreviations: CITED1, CREB-binding protein/ p300-interacting transactivator with Asp/Glu-rich C-terminal domain 1; CK19, cytokeratin 19; FA, follicular adenoma; FC, follicular carcinoma; FTUMP, follicular tumor of undetermined malignant potential; FVPTC, follicular variant of papillary thyroid carcinoma; Galectin-3, galactoside-binding lectin soluble 3; HBME1, Hector Battifora mesothelial 1; IMP3, insulin-like growth factor II messenger RNA binding protein 3; PTC, papillary thyroid carcinoma.
(93-99) Among the variety of biomarkers reported in the literature, Hector Battifora mesothelial-1 (HBME-1), galectin-3 (GAL3), cytokeratin 19 (CK19), Cbp/p300-interacting transactivator with Glu/Asp-rich carboxy-terminal domain, 1 (CITED1
), and TPO are most promising.
Heat-induced antigen retrieval was carried out for CITED1
(pH 6.1, steaming for 30 minutes at 94[degrees]C; Target Retrieval Solution, Dako).