Skip to main content

Immunohistochemical Evaluation of Biomarkers in Prostatic and Colorectal Neoplasia

Principles and Guidelines

  • Protocol
Tumor Marker Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 14))

Abstract

In Chapter 10 of thts volume and in prior publications (1,2), the matertals and methods used in immunohtstochemical techniques are described, and factors that affect the immunohistochemtcal detection of biomarker expression are discussed In this chapter, our semtquantbiattve method of evaluation of btomarker expression is explained. Also, the use of nnrnunohistochemtstry (IHC) to characterize btomarker expression in neoplastic lesions of the prostate and colorectum is dtscussed. The use of btomarkers in the study of neoplasia has expanded in recent years as new uses for biomarkers have been identified. For example, our laboratory has studied biomarkers to characterize tumorigenesis, support novel therapies, aid in diagnoses, predict aggressive tumor subtypes, and monbior the response of oral dysplasta to chemoprevention wbih retmoids. In studying oral neoplasia, we found that the expression of transforming growth factor α (TGFα) was increased in leukoplakia and that treatment wbih 13-cis-retmotc acid caused complete or partial resolutton of leukoplakia and concombiant decreased expression of TGFα (3). Thus, TGFα could be considered a candidate surrogate intermediate endpoint btomarker for chemoprevention studtes of oral dysplasia. Similarly, we have used the evaluation of biomarker expression in tissues to qualify patients wbih tumors of the prostate, colorectum, breast, ovary, and lung for specrfic types of immuno-therapy and for immunization wbih tumor vaccines (4). We also have studied biomarker expression in order to characterize the putative preinvasive neoplastic lesion of the prostate, prostatic intraepbihelial neoplasia (PIN), and to relate PIN to invasive prostatic adenocarcinomas (58). Determination of biomarker expression in colorectal tumors has been useful in identifying racial differences in these tumors as well as differences in histopathological tumor types (9). In this chapter, we demonstrate the advantages of semiquantbiatively evaluating biomarker expression in neoplastic lesions of the prostate and colorectum.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Grizzle W. E., Myers R. B, Arnold M. M., and Snvastava S. (1994) Evaluation of blomarkers in breast and prostate cancer J Cell Blochem 19(Suppl.), 259–266

    CAS  Google Scholar 

  2. Grizzle W E., Myers R B, and Oelschlager D K. (1995) Prognostic blomarkers in breast cancer: factors affecting nnmunohlstochemlcal evaluation Breast 1, 243–250

    Article  Google Scholar 

  3. Beenken S., Huang P., Sellers M., Listmsky C., Stockard C, Hubbard W., Wheeler R., and Grizzle W. E (1994) Retinoid modulation of biomarkers in oral leukoplakta/dysplasta J Cell Biochem 19(Suppl.), 270–277

    CAS  Google Scholar 

  4. Myers R B, Meredbth R F, Schlom J, LoBuglio A F, Bueschen A L, Wheeler R. H, Stockard C. R., and Grizzle W. E. (1994) Tumor associated glycoprotem-72 is highly expressed in prostatic adenocarcinomas. J Urol 152, 243–246

    PubMed  CAS  Google Scholar 

  5. Myers R B., Srivastava S., Oelschlager D. K, and Grizzle W E. (1994) Expression of p160erbB-3 and p185erbB-2 in prostatic intraepbihebal neoplasia and prostatic adenocarcmoma. J Natl Cancer Inst 86, 1140–1145

    Article  PubMed  CAS  Google Scholar 

  6. Myers R B., Schlom J, Srivastava S, and Grizzle W. E (1995) Expresston of tumor associated glycoprotem-72 in prostatic intraeptthelial neoplasta and prostatic adenocarcmoma. Modern Pathol 8, 260–265

    CAS  Google Scholar 

  7. Myers R. B, Srivastava S., and Grizzle W E (1995) Lewis Y antigen as detected by the monoclonal antibody BR96 is expressed strongly in prostatic adenocarcmoma J Ural 153, 1572–1574

    Article  CAS  Google Scholar 

  8. Myers R B., Srtvastava S, Oelschlager D K, Brown D, and Grizzle W E (1996) Expression of nm23-H1 in prostatic mtraeptthebal neoplasia and prostatic adenocarcromia. Hum Pathol 27, 1021–1024

    Article  PubMed  CAS  Google Scholar 

  9. Manne U., Myers R. B, Moron C., Srivastava S., and Grizzle W E (1996) Racial distribution of p53 mutations in colorectal adenocarcinoma Modern Pathol 1, 62A

    Google Scholar 

  10. De Potter C. R, Quatacker J, Maertens G, Van Daele S, Pauvels C, Verhofstede C, Eechaute W, and Roels H (1989) The subcellular localizatton of the neu protem in human normal and neoplastic cells Int J Cancer 44, 969–974.

    Article  PubMed  Google Scholar 

  11. Sadasivan R, Morgan R., Jennings S., Austenfeld M., Van Veldhutzen P., Stephens R, and Noble M (1993) Overexpression of HER-2/neu may be an indicator of poor prognosis in prostattc cancer J Ural 150, 126–131

    CAS  Google Scholar 

  12. Veltri R W, Parfin A. W., Epstein J P, Marley G. M, Miller G M., Singer D S., Patton K. P, Criley S R., and Coffey D S (1994) Quantative nuclear morphometry, Markovian texture descriptors and DNA content captured on a CAS-200 image analysts system, combined wbih PCNA and HER-2-NEU immunohtstochemtstry for predtctton of prostate cancer progression. J Cell Blochem 19(Suppl.), 249–258

    CAS  Google Scholar 

  13. Thor A. D, Moore D H., Edgerton S M, Kawasaki E S, Remhouse E, Lynch H. T, Marcus J N, Schwartz L, Chen L C, and Mayall B H (1992) Accumulation of p53 tumor suppressor gene protein an independent marker of prognosis in breast cancers J Nat1 Cancer Inst 84, 845–855

    Article  CAS  Google Scholar 

  14. Lohman D L, Ruhri Ch, Schmttt M, Graeff H, and Hofler H. (1993) Accumulation of p53 protein as an indicator for p53 gene mutation in breast cancer Occurrence of false-posbiives and false negatives Diagn. Mol Pathol 2, 36–41

    Google Scholar 

  15. Barnes D. M, Dublin E A., Fisher C. J., Levison D. A., and Mtllts R R (1993) Immunohistochemtcal detection of p53 protein in mammary carcmoma. an important new independent indicator of prognosis? Hum Pathol. 24, 469–476.

    Article  PubMed  CAS  Google Scholar 

  16. Myers R B, Oelschlager D, Srivastava S., and Grizzle W. E (1994) Accumulatton of the p53 protein occurs more frequently in metastatic than in localized prostatic adenocarcinomas Prostate 25, 243–248

    Article  PubMed  CAS  Google Scholar 

  17. Vtsakorpr T, Kalhomemr O. P., Herkkmen A., Korvula T., and Isola J. (1992) Small subgroup of aggressive highly proliferative prostatic carcinomas defined by p53 accumulatton. J Natl Cancer Inst 84, 883–887

    Article  Google Scholar 

  18. Navone N M, Troncoso P, Prsters L L, Goodrow T. L, Palmer J L., Nichols W W, Von Eschenbach A. C., and Conti C. J. (1993) p53 protein accumulation and gene mutation in progression of human prostate carcinoma J Natl Cancer Inst 85, 1657–1669

    Article  PubMed  CAS  Google Scholar 

  19. Yamamoto T. M., Ikawa S, Akryama T, Semba K., Nomura N., Mlyajima N, Sabio T, and Toyoshrma K (1986) Similarbiy of protein encoded by the human c-erbB-2 gene to eprdermal growth factor receptor Nature 319, 230–234

    Article  PubMed  CAS  Google Scholar 

  20. Slamon D J, Clark G. M, Wong S G, Levm W J., Ullrrch A., and McGune W L (1987) Human breast cancer correlation of relapse and survrval wbih amphfication of the HER2lneu oncogene. Science 235, 177–182

    Article  PubMed  CAS  Google Scholar 

  21. Slamon D J., Godolphm W., Jones L. A., Holt J. A., Wong S. G., Kebih D. E., Levm W. J., Stuart S G, Udove J., Ullrrch A., and Press M. F. (1989) Studies of the HER2/neu proto-oncogene in human breast and ovarian cancer. Science 244, 707–712

    Article  PubMed  CAS  Google Scholar 

  22. Hennessy C, Henry J A., May F E., Wesfly B R, Angus B., and Lennard T. W. (1991) Expression of the antrmetastatic gene in human breast cancer an assocratton wbih good prognosis J Natl Cancer Inst 83, 28l–285

    Google Scholar 

  23. Florenes V A., Aamdal S, Myklebost O., Maelandsmo G M, Bruland O S., and Fodstad O (1992) Levels of nm23 messenger RNA in metastatrc malignant melanomas Cancer Res 52, 6088–6091

    PubMed  CAS  Google Scholar 

  24. Leone A, Seeger R C, Hong C M, Hu Y Y, Arboleda M. J., Brodeur G M, Stram D, Slamon D J., and Steeg P. S (1993) Evidence for nm23 RNA overexpression, DNA amplification and mutation in aggressive chrldhood neuro-blastomas Oncogene 8, 855–865

    PubMed  CAS  Google Scholar 

  25. Engel M, Thersinger B., Seth T, Seltz G, Huwer li., Zang K. D., Welter C, and Dooley S. (1993) High levels of nm23-Hl and nm23-H2 messenger RNA in human squamous-cell lung carcmoma are associated wbih poor differentiation and advanced tumor stages. Int J Cancer 55, 375–379

    Article  PubMed  CAS  Google Scholar 

  26. Duller L., Kassel J, Nelson C E, Gryka M. A., Lttwak G, Gebhardt M, Ozturk M, Baker S J, and Vogelstem B (1990) p53 functions as a cell cycle control protein in osteosarcomas. Mol Cell Blol. 10, 5772–5781

    Google Scholar 

  27. Yomsch-Rovach E, Resmtzky D, Lotem J, Sachs L, Klmchi A, and Oren M (1991) Wild-type p53 Induces apoptosis of myelord leukaemrc cells that is inhibbied by interleukin-6 Nature 352, 345–347.

    Article  Google Scholar 

  28. Kobayasht M., Watanabe H, AJioka Y., Yoshrda M, Hbiomr J., and Asakura H. (1995) Correlatron of p53 protein expression wbih apoptotrc mcrdence in colorectal neoplasra. Virchows Archiv 427, 27–32

    Google Scholar 

  29. Vaux D L, Cory S, and Adams T M. (1988) bcl-2 promotes the survival of haemopotettc cells and cooperates wbih c-myc to immortahze pre-b cells. Nature 335, 440–442

    Article  PubMed  CAS  Google Scholar 

  30. Clarke A. R., Purdie C A., Harrison D J., Morns R G, Bird C C, Hooper M L, and Wylhe A H. (1993) Thymocytic apoptosts induced by p53-dependent and independent pathways Nature 362, 849–852

    Article  PubMed  CAS  Google Scholar 

  31. Lotera J and Sachs L (1993) Regulation by bcl-2, c-myc, and p53 of suscepttbihty to mduction of apoptosis by heat shock and cancer chemotherapy compounds in differentiation-competent and defective myelord leukemic cells Cell Growth Differ 4, 41–47

    Google Scholar 

  32. Chang E H, Ptrollo K F., Zou Z. Q, Cheung H. Y, Lawler E. L., Garner R, Whbie E, Bernstein W B, Fraumenr J F, and Blattnei W A (1987) Oncogenes in radioresistant noncancerous skin fibroblasts from a cancer-prone family Science 237, 1036–1039

    Article  PubMed  CAS  Google Scholar 

  33. Stlvestrmt R., Venerom S., Datdone M. G., Benmt E, Borachi P., Mazzetti M, Di Fronzo G, Rilke F, and Veronest U (1994) The bcl-2 protem. a prognosttc mdrcator strongly related to p53 protem in lymph node-negative breast cancer pattents. J Natl Cancer Inst 86, 499–504

    Article  Google Scholar 

  34. Gorczyca W., Marktewski M, Kram A, Tuztak T, and Domagala W Immuno-htstochemtcal analysts of bcl-2 and p53 expression in breast carcinomas their correlatton wbih K1-67 growth fraction Virchows Archiv 426, 229–233

    Google Scholar 

  35. Henriksen R, Wtlander E, and Oberg K (1995) Expression and prognosttc signrficance of bcl-2 in ovarian tumors Br J Cancer 72, 1324–1329

    Article  PubMed  CAS  Google Scholar 

  36. Herold J. J. O, Eliopoulos A G., Warwick J, Ntedobttek G., Young L. S, and Kerr D J (1996) The prognostic sigtuficance and p53 expression in ovarian car-cinoma Cancer Res 56, 2178–2184

    Google Scholar 

  37. McDonnell T J, Troncoso P., Brisbay S. M, Lagothetrs C., Chung L. W K, Hsieh J-T, Tu S-M, and Campbell M L. (1992) Expression of the proto-on-cogene bcl-2 in the prostate and bis assoctatton wbih emergence of androgen mdependent prostate cancer Cancer Res 52, 6940–6944

    PubMed  CAS  Google Scholar 

  38. Hague A, Moorghen M., Hicks D., Chapman M, and Paraskeva C (1994) bcl-2 expressron in human colorectal adenocarcinomas and carcinomas Onco-gene 9, 3367–3370.

    CAS  Google Scholar 

  39. Bosari S., Moneghmi L., Graziam D, Lee A K, Murray J J, Coggi G, and Vtale G (1995) bcl-2 oncoprotem in colorectal hyperplastic polyps, adenomas, and adenocarcinomas Hum Pathol. 26, 534–540

    Article  PubMed  CAS  Google Scholar 

  40. Manne U., Myers R B., Moron C., Poczantek R B., Dullard S, Wetss H, Brown D., Srtvastava S., and Grizzle W. E. (1996) Prognosttc stgmficance of Bcl-2 expression and p53 nuclear accumulatton in colorectal adenocarcmoma. Int J Cancer 74, 346–358

    Article  Google Scholar 

  41. Fontanini G, Vignatt S., Btgmi D, Musst A., Lucchi M, Angelettt C A, Basolo F., and Bevtlacqua G. (1995) bcl-2 proteins a prognosttc factor inversely correlated to p53 in non-small-cell lung cancer Br J Cancer 71, 1003–1007.

    Article  PubMed  CAS  Google Scholar 

  42. Roncalh V, Grimelms L, Graziam D., Wtlander E, Johansson H, Bergholm U., and Coggi G. (1995) Prognostic value of bcl-2 immunoreactivbiy in medullary thyroid carcmoma Hum Pathol 26 945–950

    Article  Google Scholar 

  43. Pezella F, Morrison H, Jones M., Gatter K. C, Lane D, Hams A. L, and Mason D Y (1993) Immunohistochemical detection of p53 and bcl-2 protems in non-Hodgkin’s lymphoma Histopathology 22, 39–44.

    Article  Google Scholar 

  44. Wynford-Thomas D (1992) p53 in tumor pathology-can we trust immunocyto-chemistry? J Pathol 166, 329–330.

    Article  PubMed  CAS  Google Scholar 

  45. Allred C D, Clark M G., Elledge R., Fuqua S. A, Brown R. W., Chamness G C, Osborne C K, and McGune W L (1993) Association of p53 protein expression with tumor cell prohferatton rate and clinical outcome in node-negative breast cancer. J. Natl Cancer Inst 85, 200–206

    Article  PubMed  CAS  Google Scholar 

  46. Nogucht M., Maezawa N., Nakawishi Y., Matsuno Y., Shimosata Y., and Htrohasht S. (1993) Apphcatton of the p53 gene mutation pattern for differential diagnosts of primary versus metastatic lung carcinomas. Diagn Mol. Pathol 2, 29–35.

    Google Scholar 

  47. Onta M, Iwahana H, Kanazawa H, Hayashi K., and Sektya T (1989) Detection of polymorphisms of human DNA by gel electrophoresis of single-strand conformation polymorphtsms Proc Natl Acad Sci. USA 86, 2776–2780.

    Google Scholar 

  48. Murakamt Y, Hayashl K., Htrohasht S., and Sekiya T. (1991) Aberrations of the tumor suppressor-p53 and retmoblastoma genes in human hepatocellular car cinoma. Cancer Res 51, 552–5525.

    Google Scholar 

  49. Moll U M, Riou G, and Levine A J (1992) Two distinct mechanisms alter p53 in breast cancer mutation and nuclear exclusion Proc. Natl Acad Sci USA 89, 7262–7266.

    Article  PubMed  CAS  Google Scholar 

  50. Kennedy S M., Macgeogh C, Jaffe R, and Spurr N. K (1994) Overexpression of the oncoprotem p53 in prtmary hepatic tumors of childhood does not correlate with gene mutations Hum Pathol 25, 438.

    Article  PubMed  CAS  Google Scholar 

  51. Xu L., Chen Y-T, Huvos A. G., Zlotolow I M, Retttg W., Old L J., and Ganin Chess P (1994) Overexpression of p53 in squamous cell carcinomas of head and neck without apparent gene mutations. Diagn Mol Pathol. 3, 83

    Article  PubMed  CAS  Google Scholar 

  52. Heimdal K, Lothe R A., Lystad S, Holm R, Fossa S. D., and Borresen A L (1993) No germline Tp53 mutations detected in familial and bilateral testtcular cancer Genes Chrom Cancer (Philadelphia) 6, 92–97

    Article  CAS  Google Scholar 

  53. Peng H Q, Hogg D., Malkm D, Bailey D, Gallie B. L, Bulbul M, Jewell M, Buchanan J, and Goss P E. (1993) Mutations of p53 gene do not occur in testis cancer Cancer Res 53, 3574–3578

    PubMed  CAS  Google Scholar 

  54. Battifora H and Kopmski M (1986) The influence of protease digestion and duration of fixation on the immunostaining of keratms: a comparison of formalm and ethanol fixation. J Hlstochem Cytochem 34, 1095–1100

    Article  CAS  Google Scholar 

  55. Ordonez M A, Manning J T, and Brooks T. E. (1988) Effect of trypsmization on the immunostaining of formalin-fixed paraffin embedded tissues. Am J Surg Pathol 12, 121–129.

    Article  PubMed  CAS  Google Scholar 

  56. Andrade R E, Hagen K. A, and Swanson P E. (1988) The use of proteolysis with ficm for mununostaining of paraffin sections Am J Clin Pathol 90, 33–39.

    PubMed  CAS  Google Scholar 

  57. Sht S.-R., Key M E, and Kalra K. L. (1991) Antigen retrieval in formalin-fixed, paraffin-embedded tissues. an enhancement method for mnnunohistochemical staining based upon microwave heating of tissue sections J Hlstochem Cytochem 39, 74l–748

    Google Scholar 

  58. Cattoretti G, Becker H M G, and Key G (1992) Monoclonal antibodies against recombinant parts of the Ki-67 antigen (MIB 1 and MIB 3) detect prohferating cells in microwave-processed formalm-fixed paraffin sections J Puthol 168, 357–363

    Article  CAS  Google Scholar 

  59. Gown A M, de Wever N., and Batttfora H. (1993) Microwave-based anttgemc unmasking. a revolutionary new techmque for routine immunohistochemistry Appl Immunohlstochem 1, 256–266

    CAS  Google Scholar 

  60. Brown R W and Chirala R (1995) Utility of microwave-cbirate antigen retrieval in diagnostic immunohistochemtstry. Modern Pathol 8, 515–520

    CAS  Google Scholar 

  61. Wood G. S. and Warnke R (1981) Suppression of endogenous avidin-binding activbiy in tissues and tts relevance to biotm-avidm detection systems. J Hutothem. Cytochem 29, 1196–1204

    Article  CAS  Google Scholar 

  62. Hsu S. M, Raine L., and Fanger H Use of avidm-biotin peroxidase complex (ABC) in unmunoperoxidase technique. a comparison between ABC and unla-beled antibody (PAP) procedures. J Hzstochem Cytochem 29, 577–580

    Google Scholar 

  63. Mazur M T, Hendrickson M R, and Kempson R L (1983) Optically clear nuclei: an alteration of endometrial eprthelium in the presence of trophoblast Am J Surg Pathol 1, 415–423

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc, Totowa, NJ

About this protocol

Cite this protocol

Grizzle, W.E., Myers, R.B., Manne, U., Srivastava, S. (1998). Immunohistochemical Evaluation of Biomarkers in Prostatic and Colorectal Neoplasia. In: Hanausek, M., Walaszek, Z. (eds) Tumor Marker Protocols. Methods in Molecular Medicine™, vol 14. Springer, Totowa, NJ. https://doi.org/10.1385/0-89603-380-5:143

Download citation

  • DOI: https://doi.org/10.1385/0-89603-380-5:143

  • Publisher Name: Springer, Totowa, NJ

  • Print ISBN: 978-0-89603-380-1

  • Online ISBN: 978-1-59259-598-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics