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Limitations with light microscopy in the detection of colorectal cancer cells

  • Original Contributions
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Diseases of the Colon & Rectum

Abstract

PURPOSE: The failure of light microscopy to predict individual patient survival accurately in pStage I and II colorectal carcinoma can hinder planning postoperative therapy and follow-up. This study was designed and conducted in two parts to assess the influence of relative sensitivity of the light microscope on the pathologist's ability to detect malignant cells in lymph nodes. METHODS: The first part of the study examined the issue of sampling error as a fraction of the number of lymph node sections examined by asking the question, “Does increasing the number of sections (sampling) taken from the block increase tumor cell detection in a lymph node?” Three levels of five sections 4 to 5 µm thick separated by 15 to 20 µm were obtained from each of 494 blocks from 173 cases of pStage I and II colorectal carcinoma. A total of 1,721 lymph nodes were examined. Sections from each level were stained with hematoxylin and eosin and for the expression of cytokeratin. The second part of the study examined the relative sensitivity of the light microscope to detect tumor cells in a lymph node. To simulate lymph nodes, cell blocks were made that contained 106 or 107 mononuclear cells admixed with increasing numbers of SW480 tumor cells (0, 50, 102, 5×102, 103, and 5×103). Three pathologists independently examined sections from ten control and ten experimental blocks. RESULTS: Results from the first part of the study demonstrated cytokeratin-positive cells in 278 lymph nodes from 102 of 172 (59 percent) cases. These cells were identified in the first level in 177 (64 percent) as compared with the second or third level or both in 101 (36 percent) of the lymph nodes. Results from the second part of the study demonstrated an overall sensitivity of light microscopic examination of hematoxylin and eosin-stained sections to be approximately 23 percent, representing tumor cells correctly detected in 7 sections of the 30 sections containing tumor cells. The overall specificity was 87 percent or 26 sections correctly classified as lacking tumor cells of a possible 30. Immunohistochemical staining for cytokeratin expression improved sensitivity of the light microscope to detect tumor cells to 18 of 30 (60 percent) and the specificity to 30 of 30 (100 percent). CONCLUSION: This study demonstrates several sources of variability that can induce errors in pathologic staging. These include 1) inadequate section,i.e., sampling, of lymph nodes, 2) use of only hematoxylin and eosin-stained sections, 3) samples with tumor cells below the level of detection sensitivity of the light microscope, and 4) observer variability.

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References

  1. Cancer Facts, Figures—1998. Atlanta: American Cancer Society, 1998.

  2. Kennedy BJ. Aging and cancer. In: Balducci L, Layman GH, Ershler WB, eds. Geriatric Oncology. Philadelphia: JB Lippincott, 1992:1–7.

    Google Scholar 

  3. Hernanz F, Revuelta S, Redondo C, Madrazo C, Castillo J, Gomez-Fleitas M. Colorectal adenocarcinoma: quality of the assessment of lymph node metastases. Dis Colon Rectum 1994;37:373–7.

    Article  PubMed  Google Scholar 

  4. Cruciti F, Doglietto GB, Bellantone R,et al. Accurate specimen preparation and examination is mandatory to detect lymph nodes and avoid understaging in colorectal cancer. J Surg Oncol 1992;51:153–8.

    PubMed  Google Scholar 

  5. Kotanagi H, Fukuoka T, Shibata Y,et al. The size of regional lymph nodes does not correlate with the presence or absence of metastasis in lymph nodes in rectal cancer. J Surg Oncol 1993;54:252–4.

    PubMed  Google Scholar 

  6. Scott KW, Grace RH. Detection of lymph node metastases in colorectal carcinoma before and after fat clearance. Br J Surg 1989;76:1165–7.

    PubMed  Google Scholar 

  7. Haboubi NY, Clark P, Kaftan SM, Schofield PF. The importance of combining xylene clearance and immunohistochemistry in the accurate staging of colorectal carcinoma. Br J Med 1992;85:386–8.

    Google Scholar 

  8. Herrera L, Villarreal JR. Incidence of metastases from rectal adenocarcinoma in small lymph nodes detected by clearing technique. Dis Colon Rectum 1992;35:783–8.

    Article  PubMed  Google Scholar 

  9. Cutait R, Alves VA, Lopes LC,et al. Restaging of colorectal cancer based on the identification of lymph node micrometastases through immunoperoxidase staining of CEA and cytokeratins. Dis Colon Rectum 1991;34:917–20.

    Article  PubMed  Google Scholar 

  10. Jeffers MD, O'Dowd GM, Mulcahy H, Stagg M, O'Donoghue DP, Toner M. The prognostic significance of immunohistochemical detected lymph node micrometastases in colorectal carcinoma. J Pathol 1994;172:183–7.

    Article  PubMed  Google Scholar 

  11. Greenson JK, Isenhart CE, Rice R, Mojzisik C, Houchens D, Martin EW Jr. Identification of occult micrometastases in pericolic lymph nodes of Dukes' B colorectal cancer patients using monoclonal antibodies against cytokeratin and CC49; correlation with long-term survival. Cancer 1994;73:563–9.

    PubMed  Google Scholar 

  12. Makin CA, Bobrow LG, Nicholls RJ. Can immunohistology improve detection of lymph node metastases in large-bowel cancer? Dis Colon Rectum 1989;32:99–102.

    PubMed  Google Scholar 

  13. Cote RJ, Houchens DP, Hitchcock CL,et al. Intraoperative detection of occult colon cancer micrometastases using125I-radiolabeled monoclonal antibody CC49. Cancer 1996;77:613–20.

    Article  PubMed  Google Scholar 

  14. Hayashi N, Arakawa H, Nagase H,et al. Genetic diagnosis identifies occult lymph node metastases undetectable by the histopathological method. Cancer Res 1994;54:3853–6.

    PubMed  Google Scholar 

  15. Hayashi N, Ito I, Yanagisawa A,et al. Genetic diagnosis of lymph node metastasis in colorectal cancer. Lancet 1995;345:1257–9.

    Article  PubMed  Google Scholar 

  16. Mori M, Mimori K, Inoue H,et al. Detection of cancer micrometastases in lymph nodes by reverse transcriptase-polymerase chain reaction. Cancer Res 1995;55:3417–20.

    PubMed  Google Scholar 

  17. Sedmak DD, Meineke TA, Knechtges DS, Anderson J. Prognostic significance of cytokeratin-positive breast cancer metastases. Mod Pathol 1989;2:516–20.

    PubMed  Google Scholar 

  18. Ludwig Breast Cancer Study Group. Prognostic importance of occult axillary lymph node micrometastases from breast cancer. Lancet 1990;335:1565–8.

    Google Scholar 

  19. Neville AM. Breast cancer micrometastases in lymph nodes and bone marrow are prognostically important. Ann Oncol 1991;2:13–4.

    PubMed  Google Scholar 

  20. Osborne MP, Asina S, Wong GY, Old LJ, Cote RJ. Immunofluorescent monoclonal antibody detection of breast cancer in bone marrow. A study of sensitivity in a model system. Cancer Res 1989;49:2510–3.

    PubMed  Google Scholar 

  21. Molina A, Colombatti M, Bonetti F,et al. A comparative analysis of three different techniques for the detection of breast cancer cells in bone marrow. Cancer 1991;67:1033–6.

    PubMed  Google Scholar 

  22. Gross HJ, Verwer B, Houck D, Hoffman RA, Recktenwald D. Model study detecting breast cancer cells in peripheral blood mononuclear cells at frequencies as low as 10(−7). Proc Natl Acad Sci U S A 1995;92:537–41.

    PubMed  Google Scholar 

  23. Hardingham JE, Kotasek D, Farmer B,et al. Immunobead-PCR: a technique for the detection of circulating tumor cells using immunomagnetic beads and the polymerase chain reaction. Cancer Res 1993;53:3455–8.

    PubMed  Google Scholar 

  24. Gluck S, Ross AA, Layton TJ,et al. Decrease in tumor cell contamination and progenitor cell yield in leukapheresis products after consecutive cycles of chemotherapy for breast cancer treatment. Biol Blood Marrow Transplant 1997;3:316–23.

    PubMed  Google Scholar 

  25. Edelstein RA, Zietman AL, de las Morenas A,et al. Implications of prostate micrometastases in pelvic lymph nodes: an archival tissue study. Urology 1996;47:370–7.

    Article  PubMed  Google Scholar 

  26. Cochran AJ, Wen DR, Morton DL. Occult tumor cells in lymph nodes of patients with pathological stage I malignant melanoma. An immunohistochemical study. Am J Surg Pathol 1988;12:612–8.

    PubMed  Google Scholar 

  27. Wang X, Heller R, VanVoorhis N.,et al. Detection of submicroscopic lymph node metastases with polymerase chain reaction in patients with malignant melanoma. Ann Surg 1994;220:768–74.

    PubMed  Google Scholar 

  28. Sosolik RC, McGaughy VR, DeYoung BR. Anti-MOC-31: A potential addition to the pulmonary adenocarcinoma versus mesothelioma immunohistochemistry panel. Mod Pathol 1997;10:716–9.

    PubMed  Google Scholar 

  29. Gross H-J, Verwer B, Houck D, Recktenwald D. Detection of rare cells at a frequency of one per million by flow cytometry. Cytometry 1993;14:519–26.

    Article  PubMed  Google Scholar 

  30. Fidler IJ, Ellis LM. The implications of angiogenesis for the biology and therapy of cancer metastasis. Cell 1994;79:185–8.

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. From the Department of Pathology, The Ohio State University and Arthur G. James Cancer Hospital and Research Institute, USA

    Charles L. Hitchcock M.D., Ph.D.

  2. Department of Surgery, The Ohio State University and Arthur G. James Cancer Hospital and Research Institute, USA

    James Sampsel M.D., Edward W. Martin Jr. M.D. & Mark W. Arnold M.D.

  3. Biostatistics Core, The Ohio State University Comprehensive Cancer Center and Arthur G. James Cancer Hospital and Research Institute, Columbus, Ohio

    Donn C. Young Ph.D.

Authors
  1. Charles L. Hitchcock M.D., Ph.D.
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  2. James Sampsel M.D.
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  3. Donn C. Young Ph.D.
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  4. Edward W. Martin Jr. M.D.
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  5. Mark W. Arnold M.D.
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Additional information

This work was supported in part by a grant from the Neoprobe Corporation, Dublin, Ohio.

Read in part at the meeting of The American Society of Colon and Rectal Surgeons, San Antonio, Texas, May 2 to 7, 1998.

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Hitchcock, C.L., Sampsel, J., Young, D.C. et al. Limitations with light microscopy in the detection of colorectal cancer cells. Dis Colon Rectum 42, 1046–1052 (1999). https://doi.org/10.1007/BF02236701

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