Indirect immunofluorescence studies of proliferating cell nuclear antigen in nucleoli of human tumor and normal tissues

Pui Kwong Chan, Rebecca Frakes, Michael G. Brattain, Karel Smetana, Harris Busch, Eng M. Tan

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

A proliferating cell nuclear antigen (PCNA) was identified with autoantibodies from a patient with systemic lupus erythematosus. Specific antibodies were purified by affinity chromatography in which Novikoff hepatoma nucleolar proteins were conjugated to Sepharose-4B. The purified anti-PCNA antibodies produced bright nucleolar fluorescence in tumor cells as shown by indirect immunofluorescence. PCNA was found in nucleoli of human cell lines, HeLa, Hep-2, and Namalwa, and a solid human renal and a prostate carcinoma. Both strong and weak nucleolar fluorescence areas were found in the renal and prostate carcinoma indicating that there are varying degrees of proliferation among tumor cells. Two human colon carcinoma cell lines, Ω (an aggressive, fast-growing clone of human colon carcinoma cell line HCT116) and CBS [a slow-growing human colon carcinoma cell line (group 3)], with different growth rates were compared. The fast-growing colon carcinoma cells, Ω, exhibited a higher percentage of nucleolar fluorescence (28.5%) than that of the slow growing colon cells (13.6%). By enzyme-linked immunosorbent assays, the Ω cell extract had a higher PCNA antigen content (2.8-fold) than that of the CBS cell extract which, in turn, was higher than that of human liver extract. PCNA was also found in a human fetal lung fibroblast cell line (IMR-90). Very weak or negative nucleolar fluorescence was observed in several normal human tissues including liver, kidney, prostate, and cheek cells. Nucleolar fluorescence was also observed in rat Novikoff hepatoma cells. Although normal rat livers do not have PCNA nucleolar fluorescence, nuclear and nucleolar fluorescence were observed at 18 hr after partial hepatectomy.

Original languageEnglish (US)
Pages (from-to)3770-3777
Number of pages8
JournalCancer Research
Volume43
Issue number8
StatePublished - Aug 1 1983

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Proliferating Cell Nuclear Antigen
Indirect Fluorescent Antibody Technique
Fluorescence
Colon
Carcinoma
Neoplasms
Cell Line
Experimental Liver Neoplasms
Prostate
Cell Extracts
Kidney
Liver Extracts
Antibodies
Cheek
Liver
Hepatectomy
Nuclear Proteins
Affinity Chromatography
Systemic Lupus Erythematosus
Autoantibodies

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Chan, P. K., Frakes, R., Brattain, M. G., Smetana, K., Busch, H., & Tan, E. M. (1983). Indirect immunofluorescence studies of proliferating cell nuclear antigen in nucleoli of human tumor and normal tissues. Cancer Research, 43(8), 3770-3777.

Indirect immunofluorescence studies of proliferating cell nuclear antigen in nucleoli of human tumor and normal tissues. / Chan, Pui Kwong; Frakes, Rebecca; Brattain, Michael G.; Smetana, Karel; Busch, Harris; Tan, Eng M.

In: Cancer Research, Vol. 43, No. 8, 01.08.1983, p. 3770-3777.

Research output: Contribution to journalArticle

Chan, PK, Frakes, R, Brattain, MG, Smetana, K, Busch, H & Tan, EM 1983, 'Indirect immunofluorescence studies of proliferating cell nuclear antigen in nucleoli of human tumor and normal tissues', Cancer Research, vol. 43, no. 8, pp. 3770-3777.
Chan PK, Frakes R, Brattain MG, Smetana K, Busch H, Tan EM. Indirect immunofluorescence studies of proliferating cell nuclear antigen in nucleoli of human tumor and normal tissues. Cancer Research. 1983 Aug 1;43(8):3770-3777.
Chan, Pui Kwong ; Frakes, Rebecca ; Brattain, Michael G. ; Smetana, Karel ; Busch, Harris ; Tan, Eng M. / Indirect immunofluorescence studies of proliferating cell nuclear antigen in nucleoli of human tumor and normal tissues. In: Cancer Research. 1983 ; Vol. 43, No. 8. pp. 3770-3777.
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abstract = "A proliferating cell nuclear antigen (PCNA) was identified with autoantibodies from a patient with systemic lupus erythematosus. Specific antibodies were purified by affinity chromatography in which Novikoff hepatoma nucleolar proteins were conjugated to Sepharose-4B. The purified anti-PCNA antibodies produced bright nucleolar fluorescence in tumor cells as shown by indirect immunofluorescence. PCNA was found in nucleoli of human cell lines, HeLa, Hep-2, and Namalwa, and a solid human renal and a prostate carcinoma. Both strong and weak nucleolar fluorescence areas were found in the renal and prostate carcinoma indicating that there are varying degrees of proliferation among tumor cells. Two human colon carcinoma cell lines, Ω (an aggressive, fast-growing clone of human colon carcinoma cell line HCT116) and CBS [a slow-growing human colon carcinoma cell line (group 3)], with different growth rates were compared. The fast-growing colon carcinoma cells, Ω, exhibited a higher percentage of nucleolar fluorescence (28.5{\%}) than that of the slow growing colon cells (13.6{\%}). By enzyme-linked immunosorbent assays, the Ω cell extract had a higher PCNA antigen content (2.8-fold) than that of the CBS cell extract which, in turn, was higher than that of human liver extract. PCNA was also found in a human fetal lung fibroblast cell line (IMR-90). Very weak or negative nucleolar fluorescence was observed in several normal human tissues including liver, kidney, prostate, and cheek cells. Nucleolar fluorescence was also observed in rat Novikoff hepatoma cells. Although normal rat livers do not have PCNA nucleolar fluorescence, nuclear and nucleolar fluorescence were observed at 18 hr after partial hepatectomy.",
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