Efficient immortalization of luminal epithelial cells from human mammary gland by introduction of simian virus 40 large tumor antigen with a recombinant retrovirus.
When defined in terms of markers for normal cell lineages, most invasive breast cancer cells correspond to the phenotype of the common luminal epithelial cell found in the terminal ductal lobular units. Luminal epithelial cells cultured from milk, which have limited proliferative potential, have now been immortalized by introducing the gene encoding simian virus 40 large tumor (T) antigen. Infection with a recombinant retrovirus proved to be 50-100 times more efficient than calcium phosphate transfection, and of the 17 cell lines isolated, only 5 passed through a crisis period as characterized by cessation of growth. When characterized by immunohistochemical staining with monoclonal antibodies, 14 lines showed features of luminal epithelial cells and of these, 7 resembled the common luminal epithelial cell type in the profile of keratins expressed. These cells express keratins 7, 8, 18, and 19 homogeneously and do not express keratin 14 or vimentin; a polymorphic epithelial mucin produced in vivo by luminal cells is expressed heterogeneously and the pattern of fibronectin staining is punctate. Although the cell lines have a reduced requirement for added growth factors, they do not grow in agar or produce tumors in the nude mouse. When the v-Ha-ras oncogene was introduced into two of the cell lines by using a recombinant retrovirus, most of the selected clones senesced, but one entered crisis and emerged after 3 months as a tumorigenic cell line.
Bartek
J
J
Imperial Cancer Research Fund, London, United Kingdom.
Bartkova
J
J
Kyprianou
N
N
Lalani
E N
EN
Staskova
Z
Z
Shearer
M
M
Chang
S
S
Taylor-Papadimitriou
J
J
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D015139:Blotting, Southern; D001940:Breast / Q000166:cytology*; D002455:Cell Division; D002460:Cell Line; D002472:Cell Transformation, Viral; D004279:DNA, Viral / Q000235:genetics; D004847:Epithelial Cells; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D007633:Keratins / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D008895:Milk, Human / Q000166:cytology*; D015689:Oncogene Protein p21(ras) / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics
D000952:Antigens, Polyomavirus Transforming; D004279:DNA, Viral; D007633:Keratins; D015689:Oncogene Protein p21(ras)