Abstract
Measurements of cell proliferation can be used as biomarkers of preneoplastic change. In this study, two immunocytochemical methods that measure different components of the cell cycle were compared to assess cell proliferation on biopsy samples from human colonic mucosa. These methods are based on a monoclonal antibody against 5-bromo-2-deoxyuridine (BrdU), which is confined to S phase cells, and a more broad assessment of proliferation based on an antibody against proliferating cell nuclear antigen (PCNA, clone 19A2). In the PCNA assay, only strongly immunostained nuclei were included. The proliferation index was assessed in colonic mucosa from patients with no colonic disorders. Correlation between individual total proliferation indices determined by either method was significant with rs = 0.6 (p < 0.05). The mean proliferation index in the study group was 7.79% using BrdU and 7.64% using PCNA immunocytochemistry. Distribution of labelled cells within crypts was similar with respect to the two methods with a peak at the 18th and the 24th percentile in the case of BrdU and at the 23rd percentile for PCNA. Variance component analysis showed that at least two biopsy specimens should be evaluated per subject to allow a precise individual characterisation. It is concluded that PCNA (19A2) immunocytochemistry may be used as an operational marker of cell proliferation in normal colonic mucosa. A significant correlation and an agreement in the mean proliferation index between PCNA (19A2) and BrdU can only be achieved by a strictly standardised enumeration of labelled cells limited to strongly stained nuclei in the PCNA evaluation.
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Selected References
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- Battersby S., Anderson T. J. Correlation of proliferative activity in breast tissue using PCNA/cyclin. Hum Pathol. 1990 Jul;21(7):781–781. doi: 10.1016/0046-8177(90)90039-8. [DOI] [PubMed] [Google Scholar]
- Bravo R., Macdonald-Bravo H. Changes in the nuclear distribution of cyclin (PCNA) but not its synthesis depend on DNA replication. EMBO J. 1985 Mar;4(3):655–661. doi: 10.1002/j.1460-2075.1985.tb03679.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bravo R., Macdonald-Bravo H. Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J Cell Biol. 1987 Oct;105(4):1549–1554. doi: 10.1083/jcb.105.4.1549. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coltrera M. D., Gown A. M. PCNA/cyclin expression and BrdU uptake define different subpopulations in different cell lines. J Histochem Cytochem. 1991 Jan;39(1):23–30. doi: 10.1177/39.1.1670579. [DOI] [PubMed] [Google Scholar]
- Eastwood G. L., Trier J. S. Organ culture of human rectal mucosa. Gastroenterology. 1973 Mar;64(3):375–382. [PubMed] [Google Scholar]
- Fireman Z., Rozen P., Fine N., Chetrit A. Reproducibility studies and effects of bowel preparations on measurements of rectal epithelial proliferation. Cancer Lett. 1989 Apr;45(1):59–64. doi: 10.1016/0304-3835(89)90036-0. [DOI] [PubMed] [Google Scholar]
- Galand P., Degraef C. Cyclin/PCNA immunostaining as an alternative to tritiated thymidine pulse labelling for marking S phase cells in paraffin sections from animal and human tissues. Cell Tissue Kinet. 1989 Sep;22(5):383–392. doi: 10.1111/j.1365-2184.1989.tb00223.x. [DOI] [PubMed] [Google Scholar]
- Garcia R. L., Coltrera M. D., Gown A. M. Analysis of proliferative grade using anti-PCNA/cyclin monoclonal antibodies in fixed, embedded tissues. Comparison with flow cytometric analysis. Am J Pathol. 1989 Apr;134(4):733–739. [PMC free article] [PubMed] [Google Scholar]
- Gerdes J., Lemke H., Baisch H., Wacker H. H., Schwab U., Stein H. Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J Immunol. 1984 Oct;133(4):1710–1715. [PubMed] [Google Scholar]
- Gratzner H. G. Monoclonal antibody to 5-bromo- and 5-iododeoxyuridine: A new reagent for detection of DNA replication. Science. 1982 Oct 29;218(4571):474–475. doi: 10.1126/science.7123245. [DOI] [PubMed] [Google Scholar]
- Hall P. A., Levison D. A., Woods A. L., Yu C. C., Kellock D. B., Watkins J. A., Barnes D. M., Gillett C. E., Camplejohn R., Dover R. Proliferating cell nuclear antigen (PCNA) immunolocalization in paraffin sections: an index of cell proliferation with evidence of deregulated expression in some neoplasms. J Pathol. 1990 Dec;162(4):285–294. doi: 10.1002/path.1711620403. [DOI] [PubMed] [Google Scholar]
- Kurki P., Ogata K., Tan E. M. Monoclonal antibodies to proliferating cell nuclear antigen (PCNA)/cyclin as probes for proliferating cells by immunofluorescence microscopy and flow cytometry. J Immunol Methods. 1988 Apr 22;109(1):49–59. doi: 10.1016/0022-1759(88)90441-3. [DOI] [PubMed] [Google Scholar]
- Lipkin M., Blattner W. E., Fraumeni J. F., Jr, Lynch H. T., Deschner E., Winawer S. Tritiated thymidine (phi p, phi h) labeling distribution as a marker for hereditary predisposition to colon cancer. Cancer Res. 1983 Apr;43(4):1899–1904. [PubMed] [Google Scholar]
- Lippman S. M., Lee J. S., Lotan R., Hittelman W., Wargovich M. J., Hong W. K. Biomarkers as intermediate end points in chemoprevention trials. J Natl Cancer Inst. 1990 Apr 4;82(7):555–560. doi: 10.1093/jnci/82.7.555. [DOI] [PubMed] [Google Scholar]
- Ogata K., Kurki P., Celis J. E., Nakamura R. M., Tan E. M. Monoclonal antibodies to a nuclear protein (PCNA/cyclin) associated with DNA replication. Exp Cell Res. 1987 Feb;168(2):475–486. doi: 10.1016/0014-4827(87)90020-6. [DOI] [PubMed] [Google Scholar]
- Preston-Martin S., Pike M. C., Ross R. K., Jones P. A., Henderson B. E. Increased cell division as a cause of human cancer. Cancer Res. 1990 Dec 1;50(23):7415–7421. [PubMed] [Google Scholar]
- Risio M., Coverlizza S., Ferrari A., Candelaresi G. L., Rossini F. P. Immunohistochemical study of epithelial cell proliferation in hyperplastic polyps, adenomas, and adenocarcinomas of the large bowel. Gastroenterology. 1988 Apr;94(4):899–906. doi: 10.1016/0016-5085(88)90545-8. [DOI] [PubMed] [Google Scholar]
- Risio M., Lipkin M., Candelaresi G., Bertone A., Coverlizza S., Rossini F. P. Correlations between rectal mucosa cell proliferation and the clinical and pathological features of nonfamilial neoplasia of the large intestine. Cancer Res. 1991 Apr 1;51(7):1917–1921. [PubMed] [Google Scholar]
- Shamsuddin A. K., Weiss L., Phelps P. C., Trump B. F. Colon epithelium. IV. Human colon carcinogenesis. Changes in human colon mucosa adjacent to and remote from carcinomas of the colon. J Natl Cancer Inst. 1981 Feb;66(2):413–419. [PubMed] [Google Scholar]
- Wargovich M. J., Medline A., Bruce W. R. Early histopathologic events to evolution of colon cancer in C57BL/6 and CF1 mice treated with 1,2-dimethylhydrazine. J Natl Cancer Inst. 1983 Jul;71(1):125–131. [PubMed] [Google Scholar]
- Wilson R. G., Smith A. N., Bird C. C. Immunohistochemical detection of abnormal cell proliferation in colonic mucosa of subjects with polyps. J Clin Pathol. 1990 Sep;43(9):744–747. doi: 10.1136/jcp.43.9.744. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yamada K., Yoshitake K., Sato M., Ahnen D. J. Proliferating cell nuclear antigen expression in normal, preneoplastic, and neoplastic colonic epithelium of the rat. Gastroenterology. 1992 Jul;103(1):160–167. doi: 10.1016/0016-5085(92)91109-h. [DOI] [PubMed] [Google Scholar]
