Abstract
Fine needle aspiration (FNA) biopsy is the least invasive method of sampling breast cancer in vivo and provides material for breast cancer diagnosis. FNA has also been used to examine cellular markers to predict and monitor the effects of therapy. The aim of this study was to assess the accuracy of using FNA material compared with resected cancer for Western blotting studies of the p53 pathway, a key to tumour response to radiotherapy and chemotherapy. Paired samples of breast cancer FNAs collected pre-operatively and post-operatively were compared with tissue samples obtained at the time of surgical resection. Western blots were probed for p53 using the antibodies DO12 and DO1, and for levels of downstream proteins p21/WAF1 and p27. The protein extracted by FNA was sufficient for up to 5 Western blot studies. p53 expression and phosphorylation did not differ significantly pre- and post-operatively, indicating that intra-operative manipulation does not affect p53 expression or downstream activation in breast cancer. However, expression of p53, p21 and p27 varied between individual patients suggesting a range of p53 pathway activation in breast cancer. Immunohistochemistry confirmed that the cancer cells accounted for the protein expression detected on Western blots. FNA yields adequate protein for Western blotting studies and could be used as a method to monitor p53 activity in vivo before and during anti-cancer treatment possibly providing early evidence of tumour response to therapy. © 2001 Cancer Research Campaign http://www.bjcancer.com
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References
Aas T, Borresen AL, Geisler S, Smith-Sorensen B, Johnsen H, Varhaug JE, Akslen LA and Lonning PE (1996) Specific p53 gene mutations are associated with de novo resistance to doxorubicin in breast cancer patients. Nat Med 2: 811–814
Andersen TI and Borrensen AL (1995) Alterations of the TP53 gene as a potential prognostic marker in breast carcinomas. Advantages of using constant denaturant gel electrophoresis in mutation detection. Diagn Mol Path 4: 203–211
Barbareschi M, Caffo O, Doglioni C, Fina P, Marchetti A, Buttitta F, Leek R, Morelli L, Leonardi E, Bevilacqua G, Dalla Palma P and Harris AL (1996) p21WAF1 immunohistochemical expression in breast carcinoma: correlations with clinicopathological data, oestrogen receptor status, MIB1 expression, p53 gene and protein alterations and relapse-free survival. Br J Cancer 74: 208–215
Blaydes JP and Hupp TR (1998) DNA damage triggers DRB-resistant phosphorylation of human p53 at the CK2 site. Oncogene 17: 1045–1052
Blaydes JP, Craig Al, Wallace M, Ball HML, Traynor NJ, Gibbs NK and Hupp TR (2000) Synergistic activation of p53-dependent transcription by two cooperating damage recognition pathways. Oncogene 19: 3829–3839
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein dye binding. Annal Biochem 72: 248–252
Brotherick I, Shenton BK and Lennard TW (1995) Are fine needle breast aspirated representative of the underlying solid tumour?. Br J Cancer 72: 732–737
Bueso-Ramos CE, Manshouri T, Haidar MA, Yang Y, McCown P, Ordonez N, Glassman A, Sneige N and Albitar M (1996) Abnormal expression of MDM-2 in breast carcinomas. Breast Cancer Res Treat 37: 179–188
Craig AL, Blaydes JP, Burch LR, Thompson AM and Hupp TR (1999) Dephosphorylation of p53 at Ser20 after cellular exposure to low levels of non-ionizing radiation. Oncogene 18: 6305–6312
Elledge RM and Allred DC (1998) Prognostic and predictive value of p53 and p21 in breast cancer. Breast Cancer Res Treat 52: 79–98
Elledge RM, Lock-Lim S, Allred DC, Hilsenbeck SG and Cordner L (1995) p53 Mutation and Tamoxifen Resistance in Breast Cancer. Clin Cancer Res 1: 1203–1208
Flaman JM, Frebourg T, Moreau V, Charbonnier F, Martin C, Chappuis P, Sappino AP, Limacher JM, Bron L, Benhattar J, Tada M, Van Meir EG, Estreicher A and Iggo RD (1995) A simple p53 functional assay for screening cell lines, blood and tumours. Proc Natl Acad Sci 92: 3963–3967
Gudlaugsdottir S, Sigurdardottir V, Snorradottir M, Jonasson JG, Ogmundsdottir H and Eyfjord JE (2000) p53 mutation analysis in benign and malignant breast lesions: using needle rinses from fine-needle aspirations. Diagn Cytopathol 22: 268–274
Hartley MN, Tuffnell DJ, Hutton JL, Palmer M and Al-Jafari MS (1988) Fine needle aspiration cytology: an in vitro study of cell yield. Br J Surg 75: 380–381
Haupt Y, Maya R, Kazaz A and Oren M (1997) Mdm2 promotes the rapid degradation of p53. Nature 387: 296–299
Hopwood D, Moitra S, Vojtesek B, Johnston DA, Dillon JF and Hupp TR (1997) Biochemical analysis of the stress protein response in human oesophageal epithelium. Gut 41: 156–163
Jones JM, Cui XS, Medina D and Donehower LA (1999) Heterozygosity of p21/WAF1/CIP1 enhances tumour cell proliferation and cyclin D1-associated kinase activity in a murine mammary cancer model. Cell Growth Differ 10: 213–222
Kastan MB, Onyekwere O, Sidransky D, Vogelstein B and Craig RW (1991) Participation of p53 protein in the cellular response to DNA damage. Cancer Res 51: 6304–6311
Kuner R, Pollow K, Lehnert A, Pollow B, Scheler P, Krummenauer F, Casper F and Hoffmann G (2000) Needle biopsy vs. Conventional surgical biopsy – biochemical analysis of various prognostic factors. Zentralbl Gynakol 122: 160–164
Lavarino C, Corletto V, Mezzelani A, Della Torre G, Bartoli C, Riva C, Pierotti MA, Rilke F and Pilotti S (1998) Detection of TP53 mutation, loss of heterozygosity and DNA content in fine needle aspirates of breast carcinoma. Br J Cancer 77: 125–130
Linke SP, Clarkin KC, Di Leonardo A, Tsou A and Wahl GM (1996) A reversible p53-dependent GO/G1 cell cycle arrest induction by ribonucleotide depletion in the absence of detectable DNA damage. Genes Dev 10: 934–947
Lowe SW, Bodics S, McClatchey A, Remington L, Ruley HE, Fisher DE, Housman DE and Jacks T (1994) p53 status and the efficacy of cancer therapy in vivo. Science 226: 807–810
Makris A, Allred DC, Powles TJ, Dowsett M, Fernando IN, Trott PA, Ashley SE, Ormerod MG, Titley JC and Osborne CK (1997) Cytological evaluation of biological prognostic markers from primary breast carcinomas. Breast Cancer Res Treat 44: 65–74
Nizzoli R, Bozzetti C, Naldi N, Guazzi A, Gabrielli M, Michiara M, Camisa R, Barilli A and Cocconi G (2000) Comparison of the results of immunocytochemical assays for biologic variables on preoperative fine-needle aspirates and on surgical specimens of primary breast carcinomas. Cancer 90: 61–66
Pelosi G, Bresaola E, Rodella S, Manfrin E, Piubello Q, Schiavon I and Iannucci A (1994) Expression of proliferating cell nuclear antigen, Ki-67 antigen, oestrogen receptor protein and tumour suppressor p53 gene in cytological samples of breast cancer: an immunochemical study with clinical, pathobiological and histological correlations. Diagn Cytopath 11: 131–140
Rao JY, Apple SK, Hemstreet GP, Jin Y and Nieberg RK (1998) Single cell multiple biomarker analysis in archival breast fine needle aspiration specimens: quantitative fluorescence image analysis of DNA content, p53 and G-actin as breast cancer biomarkers. Cancer Epidemiological Biomarkers Prev 11: 1027–1033
Sgambato A, Zhang YJ, Arber N, Hibshoosh H, Doki Y, Ciaparrone ME, Santella RM, Cittadini A and Weinstein IB (1997) Deregulated expression of p27 (Kip1) in human breast cancers. Clin Cancer Res 3: 1879–1887
Sherr CJ and Roberts JM (1999) CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev 13: 1501–1512
Shiohara M, Koike K, Komiyama A and Koeffler HP (1997) p21WAF1 mutations and human malignancies. Leuk Lymphoma 26: 1–2, 35–41
Steele RJC, Thompson AM, Hall PA and Lane DP (1998) The p53 tumour suppressor gene. Br J Surg 85: 1460–1467
Stephen CW, Helminen P and Lane DP (1995) Characterisation of epitopes on human p53 using phage-displayed peptide libraries: insights into antibody -peptide interactions. J Mol Biol 248: 58–78
Tanaka N, Ishihara M, Lamphier MS, Nozawa H, Matsuyama T, Mak TW, Aizawa S, Tokino T, Oren M and Taniguchi T (1996) Cooperation of the tumour suppressors IRF-1 and p53 in response to DNA damage. Nature 382: 816–819
Taxin A, Tartter PI and Zappetti D (1997) Breast Cancer Diagnosis by Fine Needle Aspiration and Excisional Biopsy. Recurrence and Survival. Acta Cytolgica 41: 302–306
Vojtesek B, Dolezalova H, Lauerova L, Svitakova M, Havlis P, Kovarik J, Midgley CA and Lane DP (1995) Conformational changes in p53 analysed using new antibodies to the core DNA binding domain of the protein. Oncogene 10: 383–393
Ziyaie D, Hupp TR and Thompson AM (2000) p53 and breast cancer. The Breast 9: 239–246
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Ball, HL., Hupp, T., Ziyaie, D. et al. Differential p53 protein expression in breast cancer fine needle aspirates: the potential for in vivo monitoring. Br J Cancer 85, 1102–1105 (2001). https://doi.org/10.1054/bjoc.2001.2064
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DOI: https://doi.org/10.1054/bjoc.2001.2064
