Submitted July 27, 2007
Returned for revision August 25, 2007
Accepted October 25, 2007
Female Reproductive Tract
The MUC1 HMFG1 Glycoform Is a Precursor to the 214D4 Glycoform in the Human Uterine Epithelial Cell Line, HES
Peng Wang ,
JoAnne A. Julian ,
and
Daniel D. Carson *
* To whom correspondence should be addressed. E-mail: dcarson{at}udel.edu.
Abstract
MUC1, a type I transmembrane glycoprotein expressed on most epithelia and many cancer cells, is involved in embryo implantation and tumor progression. A series of antibodies directed against the MUC1 ectodomain have been used to study MUC1 expression in the female reproductive tract, sometimes with apparently contradictory results. In the current study, we used two monoclonal MUC1 antibodies, 214D4 and HMFG1, to study the relationship between these MUC1 glycoforms in the human uterine epithelial cell line, HES, and human endometrial extracts. In response to TNF stimulation, accumulation of the HMFG1-reactive forms preceded that of the 214D4-reactive forms. Following inhibition of protein synthesis by cycloheximide, HMFG1-reactive species were lost rapidly (T1/2 = 20 min) while there was no change in the level of the 214D4-reactive forms even after 80 min. HMFG1-reactive forms had smaller oligosaccharide chains than the 214D4 reactive forms and could not be detected on the cell surface of intact cells or in the shed (media) fraction although they were readily detected in permeabilized cells. Both 214D4- and HMFG1-reactive species were detected in human endometrial extracts throughout the cycle; however, consistent with the HES cell studies, the HMFG1-reactive species were both smaller and less abundant than the 214D4-reactive species. Consistent with this we found that HMFG1-reactive species were difficult to detect in tissue sections unless predigested with neuraminidase indicating that these structures are rapidly sialylated during synthesis. In contrast, 214D4-reactive species were robustly detected in both proliferative and secretory stages. Collectively, these studies indicate that the HMFG1-reactive glycoform is a precursor of the 214D4-reactive glycoform in HES cells and normal uterine epithelia. Therefore, discrepancies in patterns of MUC1 expression in other studies may be due to failure to account for these glycoform relationships.
Key words:
Epitope •
Glycoform •
HMFG1 •
MUC1
