Skip to main content
SpringerLink
Log in

Epithelial and fibroblast cell lines derived from a spontaneous mammary carcinoma in a MMTV/neu transgenic mouse

  • Articles
  • Cell and Tissue Models
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Summary

Female murine mammary tumor virus (MMTV)/neu transgenic mice, expressing a wild-type rat neu oncogene driven by an MMTV promoter, develop focal mammary adenocarcinomas that are pathologically very similar to human breast tumors. Two new cell lines were established from a mammary tumor that arose in a female MMTV/neu transgenic mouse. One of these lines, mammary carcinoma from Neu transgenic mouse A (MCNeuA), has an epithelial morphology, is cytokeratin positive, and expresses high levels of the neu transgene. Karyotyping and comparative genomic hybridization analyses demonstrated genomic alterations in the MCNeuA cell line. The other line, N202Fb3, has a fibroblast morphology, is cytokeratin negative, and expresses the neu transgene at a very low level. This cell line also expresses smooth muscle α-actin, suggesting that it is a myofibroblast line. The MCNeuA cell line is tumorigenic when injected into syngeneic MMTV/neu transgenic mice, with an in vivo doubling time of about 14 d. The rationale for establishing this tumor cell line was to provide a tumor transplantation system for rapidly assessing immunotherapeutic interventions before testing in the more cumbersome model of spontaneous tumor development in the MMTV/neu transgenic mice. Mice immunized with a Neu extracellular domain protein vaccine were protected against a subsequent inoculation of MCNeuA cells, indicating that this cell line will be useful for evaluating cancer vaccine strategies. This tumor cell line may also prove useful in studying the biological properties of the neu oncogene and its role in the malignant process. In addition, the tumor-derived fibroblast line may be useful for studying tumor-stromal cell interactions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Adams, J. M.; Cory, S. Transgenic models of tumor development. Science 254:1161–1167; 1991.

    Article  PubMed  CAS  Google Scholar 

  • Amici, A.; Venanzi, F. M.; Concetti, A. Genetic immunization against neu/erbB2 transgenic breast cancer. Cancer Immunol. Immunother. 47:183–190; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Baselga, J.; Tripathy, D.; Mendelsohn, J., et al. Phase II study of weekly intravenous recombinant humanized anti-p185HER2 monoclonal antibody in patients with HER2/neu-overexpressing metastatic breast cancer. J. Clin. Oncol. 14:737–744; 1996.

    PubMed  CAS  Google Scholar 

  • Bellacosa, A.; Franke, T. F.; Gonzalez-Portal, M. E., et al. Structure, expression and chromosomal mapping of c-akt: relationship to v-akt and its implications. Oncogene 8:745–754; 1993.

    PubMed  CAS  Google Scholar 

  • Berx, G.; Cleton-Jansen, A. M.; Nollet, F.; de Leeuw, W. J.; van de Vijver, M.; Corneliss, C.; van Roy, F. E-cadherin is a tumour/invasion suppressor gene mutated in human lobular breast cancers. Embo. J. 14:6107–6115; 1995.

    PubMed  CAS  Google Scholar 

  • Bieche, I.; Champeme, M. H.; Lidereau, R. A tumor suppressor gene on chromosome 1p32-pter controls the amplification of MYC family genes in breast cancer. Cancer Res. 54:4274–4276; 1994.

    PubMed  CAS  Google Scholar 

  • Bieche, I.; Champeme, M. H.; Matifas, F.; Cropp, C. S.; Callahan, R.; Lidereau, R. Two distinct regions involved in 1p deletion in human primary breast cancer. Cancer Res. 53:1990–1994; 1993.

    PubMed  CAS  Google Scholar 

  • Brenner, A. J.; Aldaz, C. M. Chromosome 9p allelic loss and p16/CDKN2 in breast cancer and evidence of p16 inactivation in immortal breast epithelial cells. Cancer Res. 55:2892–2895; 1995.

    PubMed  CAS  Google Scholar 

  • Campbell, M. J.; Esserman, L.; Byars, N. E.; Allison, A. C.; Levy, R. Idiotype vaccination against murine B cell lymphoma. Humoral and cellular requirements for the full expression of antitumor immunity. J. Immunol. 145:1029–1036; 1990.

    PubMed  CAS  Google Scholar 

  • Cardiff, R. D.; Muller, W. J. Transgenic mouse models of mammary tumorigenesis. Cancer Surv. 16:97–113; 1993.

    PubMed  CAS  Google Scholar 

  • Cefai, D.; Morrison, B. W.; Sckell, A.; Favre, L.; Balli, M.; Leunig, M.; Gimmi, C. D. Targeting HER-2/neu for active-specific immunotherapy in a mouse model of spontaneous breast cancer. Int. J. Cancer 83:393–400; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Chen, Y.; Hu, D.; Eling, D. J.; Robbins, J.; Kipps, T. J. DNA vaccines encoding full-length or truncated Neu induce protective immunity against Neu-expressing mammary tumors. Cancer Res. 58:1965–1971; 1998.

    PubMed  CAS  Google Scholar 

  • Concetti, A.; Amici, A.; Petrelli, C.; Tibaldi, A.; Provinciali, M.; Venanzi, F. M. Autoantibody to p185erbB2/neu oncoprotein by vaccination with xenogenic DNA. Cancer Immunol. Immunother. 43:307–315; 1996.

    Article  PubMed  CAS  Google Scholar 

  • Cool, M.; Jolicoeur, P. Elevated frequency of loss of heterozygosity in mammary tumors arising in mouse mammary tumor virus/neu transgenic mice. Cancer Res. 59:2438–2444; 1999.

    PubMed  CAS  Google Scholar 

  • Devereux, T. R.; Anna, C. H.; Patel, A. C.; White, C. M.; Festing, M. F.; You, M. Smad4 (homolog of human DPC4) and Smad2 (homolog of human JV18-1): candidates for murine lung tumor resistance and suppressor genes. Carcinogenesis 18:1751–1755; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Disis, M. L.; Grabstein, K. H.; Sleath, P. R.; Cheever, M. A. Generation of immunity to the HER-2/neu oncogenic protein in patients with breast and ovarian cancer using a peptide-based vaccine. Clin. Cancer Res. 5:1289–1297; 1999.

    PubMed  CAS  Google Scholar 

  • Disis, M. L.; Gralow, J. R.; Bernhard, H.; Hand, S. L.; Rubin, W. D.; Cheever, M. A. Peptide-based, but not whole protein, vaccines, elicit immunity to HER-2/neu, oncogenic self-protein. J. Immunol. 156:3151–3158; 1996.

    PubMed  CAS  Google Scholar 

  • Driouch, K.; Dorion-Bonnet, F.; Briffod, M.; Champeme, M. H.; Longy M.; Lidereau, R. Loss of heterozygosity on chromosome arm 16q in breast cancer metastases. Genes Chromosomes Cancer 19:185–191; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Edwards, B. K.; Howe, H. L.; Ries, L. A. G. Annual report to the nation on the status of cancer, 1973–1999, featuring implications of age and aging on the US cancer burden. Cancer 94:2766–2792; 2002.

    Article  PubMed  Google Scholar 

  • Esserman, L. J.; Lopez, T.; Montes, R.; Bald, L. N.; Fendly, B. M.; Campbell, M. J. Vaccination with the extracellular domain of p185neu prevents mammary tumor development in neu transgenic mice. Cancer Immunol. Immunother. 47:337–342; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Fendly, B. M.; Kotts, C.; Vetterlein, D., et al. The extracellular domain of HER2/neu is a potential immunogen for active specific immunotherapy of breast cancer. J. Biol. Response Modif. 9:449–455; 1990.

    CAS  Google Scholar 

  • Foster, B. A.; Gingrich, J. R.; Kwon, E. D.; Madias, C.; Greenberg, N. M. Characterization of prostatic epithelial cell lines derived from transgenic adenocarcinoma of the mouse prostate (TRAMP) model. Cancer Res. 57:3325–3330; 1997.

    PubMed  CAS  Google Scholar 

  • Freshney, R. I. Culture of animal cells: a manual of basic techniques. New York: John Wiley & Sons; 1994:166–169.

    Google Scholar 

  • Geradts, J.; Wilson, P. A. High frequency of aberrant p16(INK4A) expression in human breast cancer. Am. J. Pathol. 149:15–20; 1996.

    PubMed  CAS  Google Scholar 

  • Guy, C. T.; Webster, M. A.; Schaller, M.; Parsons, T. J.; Cardiff, R. D.; Muller, W. J. Expression of the neu protooncogene in the mammary epithelium of transgenic mice induces metastatic disease. Proc. Natl. Acad. Sci. USA 89:10578–10582; 1992.

    Article  PubMed  CAS  Google Scholar 

  • Habuchi, T.; Luscombe, M.; Elder, P. A.; Knowles, M. A. Structure and methylation-based silencing of a gene (DBCCR1) within a candidate bladder cancer tumor suppressor region at 9q32-q33. Genomics 48:277–288; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Hegi, M. E.; Devereux, T. R.; Dietrich, W. F. Allelotype analysis of mouse lung carcinomas reveals frequent allelic losses on chromosome 4 and an association between allelic imbalances on chromosome 6 and K-ras activation. Cancer Res 54:6257–6264; 1994.

    PubMed  CAS  Google Scholar 

  • Hendrich, B.; Abbott, C.; McQueen, H.; Chambers, D.; Cross, S.; Bird, A. Genomic structure and chromosomal mapping of the murine and human Mbd1, Mbd2, Mbd3, and Mbd4 genes. Mamm. Genome 10:906–912; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Herman, J. G.; Jen, J.; Merlo, A.; Baylin, S. B. Hypermethylation-associated inactivation indicates a tumor suppressor role for p15INK4B. Cancer Res. 56:722–727; 1996.

    PubMed  CAS  Google Scholar 

  • Herman, J. G.; Merlo, A.; Mao, L.; Lapidus, R. G.; Issa, J. P.; Davidson, N. E.; Sidransky, D.; Baylin, S. B. Inactivation of the CDKN2/p16/MTS1 gene is frequently associated with aberrant DNA methylation in all common human cancers. Lancet Res. 55:4525–4530; 1995.

    CAS  Google Scholar 

  • Herzog, C. R.; Wiseman, R. W.; You, M. Deletion mapping of a putative tumor suppressor gene on chromosome 4 in mouse lung tumors. Cancer Res. 54:4007–4010; 1994.

    PubMed  CAS  Google Scholar 

  • Kremmidiotis, C.; Baker, E.; Crawford, J.; Eyre, H. J.; Nahmias, J.; Callen, D. F. Localization of human cadherin genes to chromosome regions exhibiting cancer-related loss of heterozygosity. Genomics 49:467–471; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Lahti, J. M.; Valentine, M.; Xiang, J., et al. Alterations in the PITSLRE protein kinase gene complex on chromosome 1p36 in childhood neuroblastoma. Nat. Genet. 7:370–375; 1994.

    Article  PubMed  CAS  Google Scholar 

  • Lazard, D.; Sastre, X.; Frid, M. G.; Glukhova, M. A.; Thiery, J. P.; Koteliansky, V. E. Expression of smooth muscle-specific proteins in myoepithelium and stromal myofibroblasts of normal and malignant human breast tissue. Proc. Natl. Acad. Sci. USA 90:999–1003; 1993.

    Article  PubMed  CAS  Google Scholar 

  • Li, R.; Yerganian, G.; Duesberg, P.; Kraemer, A.; Willer, A.; Rausch, C.; Hehlmann, R. Aneuploidy correlated 100% with chemical transformation of Chinese hamster cells. Proc. Natl. Acad. Sci. USA 94:14506–14511; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Liu, E.; Thor, A.; He, M.; Barcos, M.; Ljung, B. M.; Benz, C. The HER2 (c-erbB-2) oncogene is frequently amplified in in situ carcinomas of the breast. Oncogene 7:1027–1032; 1992.

    PubMed  CAS  Google Scholar 

  • Mosmann, T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods 65:55–63; 1983.

    Article  PubMed  CAS  Google Scholar 

  • Muller, W. J.; Sinn, E.; Pattengale, P. K.; Wallace, R.; Leder, P. Single-step induction of mammary adenocarcinoma in transgenic mice bearing the activated c-neu oncogene. Cell 54:105–115; 1988.

    Article  PubMed  CAS  Google Scholar 

  • Nagai, H.; Negrini, M.; Carter, S. L.; Gillum, D. R.; Rosenberg, A. L.; Schwartz, G. F.; Croce, C. M. Detection and cloning of a common region of loss of heterozygosity at chromosome 1p in breast cancer. Cancer Res. 55:1752–1757; 1995.

    PubMed  CAS  Google Scholar 

  • Nagata, Y.; Furugen, R.; Hiasa, A., et al. Peptides derived from a wild-type murine proto-oncogene c-erbB-2/HER2/neu can induce, CTL and tumor suppression in syngeneic hosts. J. Immunol. 159:1336–1343; 1997.

    PubMed  CAS  Google Scholar 

  • Narducci, M. G.; Virgilio, L.; Engiles, J. B., et al. The murine Tcll, oncogene: embryonic and lymphoid cell expression. Oncogene 15:919–926; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Phelan, C. M.; Larsson, C.; Baird, S., et al. The human mammary-derived growth inhibitor (MDGI) gene: genomic structure and mutation analysis in human breast tumors. Genomics 34:63–68; 1996.

    Article  PubMed  CAS  Google Scholar 

  • Quelle, D. E.; Ashmun, R. A.; Hannon, G. J., et al. Cloning and characterization of murine p16INK4a and p15INK4b genes. Oncogene 11:635–645; 1995.

    PubMed  CAS  Google Scholar 

  • Recio, J. A.; Zambrano, N.; de La Pena, L.; Powers, C.; Siwarski, D.; Huppi, K.; Notario, V. cDNA isolation, expression, and chromosomal localization of the mouse pcph proto-oncogene. Mol. Carcinog. 26:130–136; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Ritland, S. R.; Rowse, G. J.; Chang, Y.; Gendler, S. J. Loss of heterozygosity analysis in primary mammary tumors and lung metastases of MMTV-MTAg and MMTV-neu transgenic mice. Cancer Res. 57:3520–3525; 1997.

    PubMed  CAS  Google Scholar 

  • Sacco, M. G.; Gribaldo, L.; Barbieri, O., et al. Establishment and characterization of a new mammary adenocarcinoma cell line derived from MMTV neu transgenic mice. Breast Cancer Res Treat. 47:171–180; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Sheng, Z. M.; Marchetti, A.; Buttitta, F.; Champeme, M. H.; Campani, D.; Bistocchi, M.; Lidereau, R.; Callahan, R. Multiple regions of chromosome 6q affected by loss of heterozygosity in primary human breast carcinomas. Br. J. Cancer 73:144–147; 1996.

    PubMed  CAS  Google Scholar 

  • Shepard, H. M.; Lewis, G. D.; Sarup, J. C., et al. Monoclonal antibody therapy of human cancer: taking the HER2 protooncogene to the clinie. J. Clin. Immunol. 11:117–127; 1991.

    Article  PubMed  CAS  Google Scholar 

  • Siegel, P. M.; Dankort, D. L.; Hardy, W. R.; Muller, W. J. Novel activating mutations in the neu proto-oncogene involved in induction of mammary tumors. Mol. Cell. Biol. 14:7068–7077; 1994.

    PubMed  CAS  Google Scholar 

  • Siegel, P. M.; Muller, W. J. Mutations affecting conserved cysteine residues within the extracellular domain of Neu promote receptor dimerization and activation. Proc. Natl. Acad. Sci. USA 93:8878–8883; 1996.

    Article  PubMed  CAS  Google Scholar 

  • Slamon, D.; Clark, G.; Wong, S.; Levin, W.; Ullrich, A.; McGuire, W. Human breast cancer: correlation of relapse and survival with amplification of the HER2/neu oncogene. Science 235:177–182; 1987.

    Article  PubMed  CAS  Google Scholar 

  • Slamon, D. J.; Godolphin, W.; Jones, L. A., et al. Studies of the HER2/neu proto-oncogene in human breast and ovarian cancer. Science 244:707–712; 1989.

    Article  PubMed  CAS  Google Scholar 

  • Sola, B.; Simon, D.; Mattei, M. G.; Fichelson, S.; Bordereaux, D.; Tambourin, P. E.; Guenet, J. L.; Gisselbrecht, S. Fim-1, Fim-2/c-fims, and Fim-3, three common integration sites of Friend murine leukemia virus in myeloblastic leukemias, map to mouse chromosomes 13, 18, and 3, respectively. J. Virol. 62:3973–3978; 1988.

    PubMed  CAS  Google Scholar 

  • Solin, L. J.; Haffty, B.; Fourquet, A., et al. An international collaborative study: a 15-yr experience. In: Silverstein, M., ed. Ductal carcinoma in situ of the breast. Baltimore, MD: Williams and Wilkins; 1997:385–390.

    Google Scholar 

  • Tremblay, G. B.; Tremblay, A.; Copeland, N. G.; Gilbert D. J.; Jenkins, N. A.; Labrie, F.; Giguere, V. Cloning, chromosomal localization, and functional analysis of the murine estrogen receptor beta. Mol. Endocrinol. 11:353–365; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Tsukamoto, K.; Ito, N.; Yoshimoto, M.; Kasumi, F.; Akiyama, F.; Sakamoto, G.; Nakamura, Y.; Emi, M. Allelic loss on chromosome 1p is associated with progression and lymph node metastasis of primary breast carcinoma. Cancer 82:317–322; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Van Camp, G.; Coucke, P.; Speleman, F.; Van Roy N.; Beyer, E. C.; Oostra, B. A.; Willems, P. J. The gene for human gap junction protein connexin37 (GJA4) maps to chromosome 1p35.1, in the vicinity of DIS195. Genomics 30:402–403; 1995.

    PubMed  Google Scholar 

  • Van de Vijver, M. J.; Peterse, J. L.; Mooi, W. J.; Wisman, P.; Lomans, J.; Dalesio, O.; Nusse, R. Neu-protein overexpression in breast cancer. Association with comedo-type ductal carcinoma in situ and limited prognostic value in stage II breast cancer. N. Engl. J. Med. 319:1239–1245; 1988.

    Article  PubMed  Google Scholar 

  • Van Zee, K. J.; Calvano, J. E.; Bisogna, M. Hypomethylation and increased gene expression of p16INK4a in primary and metastatic breast carcinoma as compared to normal breast tissue. Oncogene 16:2723–2727; 1998.

    Article  PubMed  Google Scholar 

  • Webster, M. A.; Cardiff, R. D.; Muller, W. J. Induction of mammary epithelial hyperplasias and mammary tumors in transgenic mice expressing a murine mammary tumor virus/activated c-src fusion gene. Proc. Natl. Acad. Sci. USA 92:7849–7853; 1995.

    Article  PubMed  CAS  Google Scholar 

  • Wei, W. Z.; Shi, W. P.; Galy, A.; Lichlyter, D.; Hernandez, S.; Groner, B.; Heilbrun, L.; Jones, R. F. Protection against mammary tumor growth by vaccination with full-length, modified human ErbB-2 DNA. Int. J. Cancer 81:748–754; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Yamashita, J.; Ogawa, M.; Sakai, K. Prognostic significance of three novel biologic factors in a clinical trial of adjuvant therapy for node-negative breast cancer. Surgery 117:601–608; 1995.

    Article  PubMed  CAS  Google Scholar 

  • Yamashita, S.; Yamashita, J.; Ogawa, M. Overexpression of group II phospholipase A2 in human breast cancer tissues in closely associated with their malignant potency. Br. J. Cancer 69:1166–1170; 1994.

    PubMed  CAS  Google Scholar 

  • Zhuang, S. M.; Eklund, I. K.; Cochran, C.; Rao, G. N.; Wiseman, R. W.; Soderkvist, P. Allelotype analysis of 2′,3′-dideoxycytidine- and 1,3-butadiene-induced lymphomas in B6C3F1 mice. Cancer Res. 56:3338–3343; 1996.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Surgery, University of California, 94115, San Francisco, California

    Michael J. Campbell, Wendy S. Wollish, Margaret Lobo & Laura J. Esserman

Authors
  1. Michael J. Campbell
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wendy S. Wollish
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Margaret Lobo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Laura J. Esserman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michael J. Campbell.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Campbell, M.J., Wollish, W.S., Lobo, M. et al. Epithelial and fibroblast cell lines derived from a spontaneous mammary carcinoma in a MMTV/neu transgenic mouse. In Vitro Cell.Dev.Biol.-Animal 38, 326–333 (2002). https://doi.org/10.1290/1071-2690(2002)038<0326:EAFCLD>2.0.CO;2

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1290/1071-2690(2002)038<0326:EAFCLD>2.0.CO;2

Key words

Search

Navigation