Cloning of partial cDNA encoding differentiation and tumor-associated mucin glycoproteins expressed by human mammary epithelium

Journal Article · Tue Sep 01 04:00:00 EDT 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.17.6060· OSTI ID:5172620

Gender, S J; Burchell, J M; Duhig, T; Lamport, D; White, R; Parker, M; Taylor-Papadimitriou, J

Human mammary epithelial cells secrete and express on their cell surfaces complex mucin glycoproteins that are developmentally regulated, tumor-associated, and highly immunogenic. Studies using monoclonal antibodies directed to these glycoproteins suggest that their molecular structures can vary with differentiation stages in the normal gland and in malignancy. To analyze the molecular nature of these glycoproteins, milk mucin was affinity-purifed and deglycosylated with hydrogen fluoride, yielding bands at 68 and 72 kDa on silver-stained gels. Polyclonal and monoclonal antibodies to the stripped core protein were developed and used to screen a lambdagt11 expression library of cDNA made from mRNA of the mammary tumor cell line MCF-7. Seven crossreacting clones were isolated, with inserts 0.1-1.8 kilobases long. RNA blot analysis, using as a probe the 1.8-kilobase insert subcloned in plasmid pUC8 (pMUC10), revealed transcripts of 4.7 and 6.4 kilobases in MCF-7 and T47D mammary tumor cells, whereas normal mammary epithelial cells from pooled milks have additional transcripts. The expression of mRNA correlates with antigen expression as determined by binding of two previously characterized anti-mucin monoclonal antibodies (HMFG-1 and HMFG-2) to seven cell lines. Restriction enzyme analysis detected a restriction fragment length polymorphism when human genomic DNA was digested with EcoRI or HinfI.

• Research Organization: Imperial Cancer Research Fund, London (England)
• OSTI ID: 5172620
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 84:17; ISSN PNASA
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
62 RADIOLOGY AND NUCLEAR MEDICINE
ANIMAL TISSUES
ANIMALS
ANTIBODIES
AUTORADIOGRAPHY
BODY
CARBOHYDRATES
CELL DIFFERENTIATION
CLONING
DNA
DNA-CLONING
ELECTROPHORESIS
EPITHELIUM
GLANDS
GLUCOPROTEINS
HYBRIDIZATION
MAMMALS
MAMMARY GLANDS
MAN
MESSENGER-RNA
MOLECULAR STRUCTURE
MONOCLONAL ANTIBODIES
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PRECIPITATION
PRIMATES
PROTEINS
RECOMBINANT DNA
RNA
SACCHARIDES
SEPARATION PROCESSES
TISSUES
VERTEBRATES