Summary
Apoptotic markers include proteins from the intrinsic and extrinsic pathways. These cascades include both pro-apoptotic and anti-apoptotic elements. The expression levels of these elements can be assessed by immunohistochemistry (IHC) and can indicate general trends in pro- versus anti-apoptotic tendencies of the cells. IHC is particularly useful when studying large cohorts of paraffin-embedded specimens. Advances in tissue microarray (TMA) technology have facilitated evaluation of large cohorts of specimens, as cores from hundreds of patients can be represented on a single glass slide and stained in a uniform fashion. In this chapter, we discuss construction and staining methods of TMAs and present examples of assessment of apoptotic marker expression in malignant and benign cells using a novel method of automated, quantitative analysis of in situ protein expression.
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References
Hanahan, D., and Weinberg, R.A. (2000) The hallmarks of cancer. Cell 100, 57–70.
Hengartner, M.O. (2000) The biochemistry of apoptosis. Nature 407, 770–776.
Zhang, X.D., Borrow, J.M., Zhang, X.Y., Nguyen, T., and Hersey, P. (2003) Activation of ERK1/2 protects melanoma cells from TRAIL-induced apoptosis by inhibiting Smac/DIABLO release from mitochondria. Oncogene 22, 2869–2881.
Wan, W.H., Fortuna, M.B., and Furmanski, P.A. (1987) rapid and efficient method for testing immunohistochemical reactivity of monoclonal antibodies against multiple tissue samples simultaneously. J Immunol Methods 103, 121–129.
Kononen, J., Bubendorf, L., Kallioniemi, A., Barlund, M., Schraml, P., Leighton, S., Torhorst, J., Mihatsch, M.J., Sauter, G., and Kallioniemi, O.P. (1998) Tissue microarrays for high-throughput molecular profiling of tumor specimens. Nat Med 4, 844–847.
Bova, G.S., Parmigiani, G., Epstein, J.I., Wheeler, T., Mucci, N.R., and Rubin, M.A. (2001) Web-based tissue microarray image data analysis: initial validation testing through prostate cancer Gleason grading. Hum Pathol 32, 417–427.
Camp, R.L., Charette, L.A., and Rimm, D.L. (2000) Validation of tissue microarray technology in breast carcinoma. Lab Invest 80, 1943–1949.
Hoos, A., Urist, M.J., Stojadinovic, A., Mastorides, S., Dudas, M.E., Leung, D.H., Kuo, D., Brennan, M.F., Lewis, J.J., and Cordon-Cardo, C. (2001) Validation of tissue microarrays for immunohistochemical profiling of cancer specimens using the example of human fibroblastic tumors. Am J Pathol 158, 1245–1251.
Giltnane, J.M., and Rimm, D.L. (2004) Technology insight: identification of biomarkers with tissue microarray technology. Nat Clin Pract Oncol 1, 104–111.
DiVito, K.A., Charette, L.A., Rimm, D.L., and Camp, R.L. (2004) Long-term preservation of antigenicity on tissue microarrays. Lab Invest 84, 1071–1078.
Kluger, H.M., DiVito, K., Berger, A.J., Halaban, R., Ariyan, S., Camp, R.L., and Rimm, D.L. (2004) Her2/neu is not a commonly expressed therapeutic target in melanoma. Melanoma Res 14, 207–210.
Divito, K.A., Berger, A.J., Camp, R.L., Dolled-Filhart, M., Rimm, D.L., and Kluger, H.M. (2004) Automated quantitative analysis of tissue microarrays reveals an association between high Bcl-2 expression and improved outcome in melanoma. Cancer Res. 64, 8773–8777.
McCarthy, M.M., Sznol, M., DiVito, K.A., Camp, R.L., Rimm, D.L., and Kluger, H.M. (2005) Evaluating the expression and prognostic value of TRAIL-R1 and TRAIL-R2 in breast cancer. Clin Cancer Res 11, 5188–5194.
Camp, R.L., Chung, G.G., and Rimm, D.L. (2002) Automated subcellular localization and quantification of protein expression in tissue microarrays. Nat Med 8, 1323–1327.
Camp, R.L., Dolled-Filhart, M., King, B.L., and Rimm, D.L. (2003) Quantitative analysis of breast cancer tissue microarrays shows that both high and normal levels of HER2 expression are associated with poor outcome. Cancer Res 63, 1445–1448.
Koscielny, S., Terrier, P., Spielmann, M., and Delarue, J.C. (1998) Prognostic importance of low c-erbB2 expression in breast tumors. J Natl Cancer Inst 90, 712.
Rubin, M.A., Zerkowski, M.P., Camp, R.L., Kuefer, R., Hofer, M.D., Chinnaiyan, A.M., and Rimm, D.L. (2004) Quantitative determination of expression of the prostate cancer protein alpha-methylacyl-CoA racemase using automated quantitative analysis (AQUA): a novel paradigm for automated and continuous biomarker measurements. Am J Pathol 164, 831–840.
Camp, R.L., Chung, G.G., and Rimm, D.L. (2002) Automated subcellular localization and quantification of protein expression in tissue microarrays. Nat Med 11, 1323–1327.
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Pick, E., McCarthy, M.M., Kluger, H.M. (2008). Assessing Expression of Apoptotic Markers Using Large Cohort Tissue Microarrays. In: Mor, G., Alvero, A.B. (eds) Apoptosis and Cancer. Methods in Molecular Biology™, vol 414. Humana Press. https://doi.org/10.1007/978-1-59745-339-4_8
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DOI: https://doi.org/10.1007/978-1-59745-339-4_8
Publisher Name: Humana Press
Print ISBN: 978-1-58829-457-9
Online ISBN: 978-1-59745-339-4
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