Establishment and characterization of a new cell line derived from a human primary breast carcinoma.
A new cell line, designated HDQ-P1, was successfully established from a primary ductal infiltrating mammary carcinoma by using a 3T3 feeder layer lethally irradiated to 60 Gy. The HDQ-P1 cells have been grown in culture for over 115 passages and have a doubling time of 60 hours. Characterization of the cell line was performed. This included morphology by light and transmission electron microscopy, karyotype, growth rate, telomerase expression, tumor antigen expression, xenograft implantation into nude mice, colony formation 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 confirmed by ultrastructural analysis, expression of cytokeratins, and epithelial membrane antigen. The HDQ-P1 cells possess an extensively rearranged and polyploid karyotype, with an average of 20 recurrent marker chromosomes. Scatchard analysis demonstrated that both primary tumor and HDQ-P1 cells were estrogen- and progesterone-receptor negative. The HDQ-P1 cells had the same 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 DNA sequencing showed a point mutation which yielded to a stop codon at the amino acid 213 in exon 6 of the TP53 gene. A five-fold amplification of C-MYC was observed in HDQ-P1 cells. No amplification of C-ERBB-2 and C-H-RAS genes were observed. This cell line presents unique characteristics and may prove to be a good experimental model for investigating breast cancer biology.
Cancer Research Center, Centre Hospitalier Universitaire de Québec, Division of Pathology, Department of Medical Biology, Quebec, Canada.
C S
CS
Wang
F
F
Goulet
J
J
Lavoie
R
R
Drouin
F
F
Auger
S
S
Champetier
L
L
Germain
B
B
Têtu
D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism; D018270:Carcinoma, Ductal, Breast / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques; D002455:Cell Division; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins; D005260:Female; D018628:Gene Dosage; D006801:Humans; D007621:Karyotyping; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D009154:Mutation; D009368:Neoplasm Transplantation; D012313:RNA; D011960:Receptors, Estrogen / Q000032:analysis; D011980:Receptors, Progesterone / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000235:genetics; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase