✍ Scribed by Dr. Vimla Band; Deborah Zajchowski; Göran Stenman; Cynthia C. Morton; Victoria Kulesa; James Connolly; Ruth Sager
No coin nor oath required. For personal study only.
A continuous line of human mammary tumor cells, called 2 I MT, has been established in culture from a pleural effusion of a 36-year-old woman with metastatic breast cancer. The cells are epithelial as shown by morphology and expression of keratins and are mammary tumor cells as shown by expression of the HMFG-2 antigenic determinant. The cells grow well both in DFCI-I, a partially defined medium containing pituitary extract and I % fetal bovine serum, and in a-minimum essential medium (a-MEM) supplemented with 10% serum, epidermal growth factor (EGF), insulin, and hydrocortisone. Karyotypic analysis of cells at early passage has shown the presence of rearranged (marker) chromosomes as well as aneuploidy with a net DNA content in the tetraploid range, confirmed by DNA cytofluorography, as well as double minute chromosomes in about 5% of the cells. Southern blots have revealed a 40-fold amplification of the ERBB2 gene and a 50-fold overexpression of its mRNA. The amplification of ERBB2 DNA was localized by in situ hybridization t o one of the marker chromosomes but not t o the double minutes. It is inferred, therefore, that at least two genes have been amplified in these cells.
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