A method to differentiate between thyroglobulin derived from normal thyroid tissue and from thyroid carcinoma based on analysis of reactivity to lectins.
OBJECTIVE
The composition of sugar chains on thyroglobulin (Tg) produced in thyroid carcinoma cells (C-Tg) is different from Tg produced in normal thyroid tissues (N-Tg). In this study, we designed a new method for detecting Tg derived from thyroid carcinoma based on the differences between C-Tg and N-Tg in the reactivity with lectins.


METHODS
Thyroglobulin preparations obtained from various thyroid tissues were incubated with lectins, and the amount of lectin-unbound Tg (ub-Tg) in the supernatant relative to Tg untreated with lectin was determined by enzyme-linked immunosorbent assay and expressed as ub-Tg(%). In addition, to study further the differences in glycosylation between C-Tg and N-Tg, concanavalin A binding to Tg digested with Staphylococcus aureus V8 protease was analyzed on nitrocellulose membrane after Western blotting.


RESULTS
The ub-Tg(%) in C-Tg from papillary carcinoma was significantly higher than in Tg from Graves' disease, benign goiter, and normal thyroid tissue for both concanavalin A and ricinus communis agglutinin-120. Concanavalin A did not appear to bind to Tg from papillary carcinoma after V8 treatment by Western blot analysis. The ub-Tg(%) in Tg from follicular adenoma was significantly higher than C-Tg from follicular carcinoma, whereas there were no differences in ub-Tg(%) between follicular carcinoma and normal thyroid tissue in concanavalin A treatment.


CONCLUSIONS
These results suggest our new methods can distinguish both between C-Tg from papillary carcinoma and N-Tg, and between follicular carcinoma and follicular adenoma in thyroid tissue specimens. Thus, this type of analysis may be applicable to differentiate C-Tg from N-Tg in thyroid aspirates for the adjunctive cytodiagnosis of thyroid carcinoma.
Maruyama
M
M
Department of Surgery, Shinshu University School of Medicine, Asahi, Matsumoto, Japan.
Kato
R
R
Kobayashi
S
S
Kasuga
Y
Y
D015415:Biomarkers / Q000032:analysis; D015153:Blotting, Western; D003208:Concanavalin A / Q000378:metabolism; D006651:Histocytochemistry / Q000379:methods; D006801:Humans; D037102:Lectins / Q000187:drug effects / Q000378:metabolism*; D037121:Plant Lectins; D012680:Sensitivity and Specificity; D013954:Thyroglobulin / Q000302:isolation & purification / Q000378:metabolism*; D013959:Thyroid Diseases / Q000378:metabolism; D013961:Thyroid Gland / Q000378:metabolism*; D013964:Thyroid Neoplasms / Q000378:metabolism*; D014909:Wheat Germ Agglutinins / Q000378:metabolism
D015415:Biomarkers; D037102:Lectins; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A; D013954:Thyroglobulin