Establishment and characterization of a new cell line derived from a humanprimary breast carcinoma

A new cell line, designated HDQ-P1, was successfully established from a pri mary ductal infiltrating mammary carcinoma by using a 3T3 feeder layer leth ally irradiated to 60 Gy. The HDQ-P1 cells have been grown in culture for o ver 125 passages and have a doubling time of 60 hours. Characterization of the cell line was performed. This included morphology by light and transmis sion electron microscopy, karyotype, growth rate, telomerase expression, tu mor antigen expression, xenograft implantation into nude mice, colony forma tion in soft agar, TP53 sequencing, and gene copy number of C-MYC, c-ERBB-2 , and C-H-RAS oncogenes. The epithelial nature of this cell line was confir med by ultrastructural analysis, expression of cytokeratins, and epithelial membrane antigen. The HDQ-P1 cells possess an extensively rearranged and p olyploid karyotype, with an average of 20 recurrent marker chromosomes. Sca tchard analysis demonstrated that both primary tumor and HDQ-P1 cells were estrogen- and progesterone-receptor negative. The HDQ-P1 cells herd the sam e expression of human telomerase reverse transcriptase as other established breast cancer cell lines such as MDA-MB-231, SK-BR-3, and MCF-7. Direct DN A sequencing showed a point mutation which yielded to a stop codon at the a mino acid 213 in exon 6 of the TP53 gene. A five-fold amplification of C-MY C was observed in HDQ-P1 cells. No amplification of C-ERBB-2 and CH-RAS gen es were observed. This cell line presents unique characteristics and may pr ove to be a good experimental model for investigating breast cancer biology . (C) 2000 Elsevier Science Inc. All rights reserved.