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...

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Veröffentlicht in:Cancer genetics and cytogenetics 2000-07, Vol.120 (1), p.58-72
Hauptverfasser: Wang, Chang Shu, Goulet, Francine, Lavoie, Josée, Drouin, Régen, Auger, François, Champetier, Serge, Germain, Lucie, Têtu, Bernard
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container_issue 1
container_start_page 58
container_title Cancer genetics and cytogenetics
container_volume 120
creator Wang, Chang Shu
Goulet, Francine
Lavoie, Josée
Drouin, Régen
Auger, François
Champetier, Serge
Germain, Lucie
Têtu, Bernard
description 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.
doi_str_mv 10.1016/S0165-4608(99)00253-8
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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. 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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. 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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.</abstract><cop>New York, NY</cop><pub>Elsevier Inc</pub><pmid>10913678</pmid><doi>10.1016/S0165-4608(99)00253-8</doi><tpages>15</tpages></addata></record>
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subjects Animal cells
Animals
Biological and medical sciences
Biotechnology
Breast Neoplasms - genetics
Breast Neoplasms - metabolism
Carcinoma, Ductal, Breast - genetics
Carcinoma, Ductal, Breast - metabolism
Cell Culture Techniques
Cell Division
DNA Mutational Analysis
DNA-Binding Proteins
Establishment of new cell lines, improvement of cultural methods, mass cultures
Eukaryotic cell cultures
Female
Fundamental and applied biological sciences. Psychology
Gene Dosage
Humans
Karyotyping
Methods. Procedures. Technologies
Mice
Mice, Nude
Middle Aged
Mutation
Neoplasm Transplantation
Receptors, Estrogen - analysis
Receptors, Progesterone - analysis
Reverse Transcriptase Polymerase Chain Reaction
RNA
Telomerase - genetics
Tumor Cells, Cultured
title Establishment and Characterization of a New Cell Line Derived from a Human Primary Breast Carcinoma
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