Characterization of mullerian inhibiting substance binding on cervical carcinoma cells demonstrated by immunocytochemistry.
Müllerian inhibiting substance (MIS) is a glycoprotein released from Sertoli cells or follicular cells of gonads, responsible for the regression of Müllerian ducts and/or Müllerian-derived tumor cells. Binding of MIS to target cells is essential for initiating regression. A human cervical carcinoma CaSki cell was examined by quantitative immunocytochemistry detected by anti-avian MIS antibody for MIS binding ability. Various treatments of WGA-peroxidase conjugate, enzyme digestion, sodium periodate or exogenous estrogen before antibody recognition were performed. It was found that the WGA partially blocked MIS binding to CaSki cell surfaces. Protease digestion of CaSki cell surfaces prior to addition of MIS or an anticervical carcinoma monoclonal antibody 1H10 (MAb 1H10), blocked the binding of MIS but not MAb 1H10 to cell surfaces. Sodium periodate and overnight exposure of CaSki cells to estrogen or diethylstilbestrol before or after fixation of the cells, did not influence MIS binding ability in vitro. MIS binding was higher on avian Müllerian duct compared with MIS binding to CaSki cells by quantitative immuno-gold labeling analysis. MAb 1H10 immuno-gold complexes binding to CaSki cells was also obtained and compared with MIS immuno-gold bindings. MIS binding site could be a polypeptide which survived sodium periodate treatment. The 'critical window' period, in which developing Müllerian ducts respond to exogenous estrogen protection from MIS regression, is possibly lost in CaSki cell.
Wang
J J
JJ
Department and Institute of Biology and Anatomy, National Defense Medical Center, Taipei, Taiwan, Republic of China.
Roffler
S R
SR
Chou
H H
HH
Yin
F Y
FY
Yin
C S
CS
D054304:Anti-Mullerian Hormone; D004054:Diethylstilbestrol / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D006023:Glycoproteins; D006131:Growth Inhibitors / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D009095:Mullerian Ducts; D010447:Peptide Hydrolases / Q000494:pharmacology; D011485:Protein Binding; D013735:Testicular Hormones / Q000032:analysis / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D002583:Uterine Cervical Neoplasms / Q000378:metabolism* / Q000473:pathology
D006023:Glycoproteins; D006131:Growth Inhibitors; D013735:Testicular Hormones; D004958:Estradiol; D004054:Diethylstilbestrol; D054304:Anti-Mullerian Hormone; D010447:Peptide Hydrolases