Cell cycle pathway perturbation in human germ cell tumors of the testis

Robert C. Flanigan, Jm Quan Jin, Bob Challenger, W Waters, Joseph M. Pyle, T. Vincent

Research output: Contribution to journalArticle

Abstract

Changes in expression of cell cycle-related proteins or their genes, and mechanisms regulating cell death (apoptosis) are now reported in many human solid tumors. Testicular germ cell tumors are unusual, in that most respond clinically to DNA damaging chemotherapeutic agents, suggesting that cell cycle and related apoptosis pathways are intact in these tumors. Since previous reports have indicated that mutations to the p53 gene are rare in these tumors, while expression of stable p53 protein is commonly seen, we have undertaken a study of cell cycle regulatory pathways to determine the defect or defects present. Paraffin-embedded tissue sections (5-6 micron) from 26 different testicular tumors have been evaluated to date, including 16 pure seminomas (stage I or II). Expression of cell cycle related proteins was determined by immunohistochemistry (IHC) using antigen retreival (TUF, PharMingen, 90° C for 30 min) and the following antibodies: anti-p53 (DO-7, DAKO), anti-PCNA (19A2, Coulter), Ki-67 (MIB-1, Immunotech), p21 (EA-10, CalBiochem). Apoptotic nuclei were evaluated using a modified TUNEL assay (Apo-BRDU, Phoenix Flow Systems). Tissues were lightly counterstained with hematoxalin. For all IHC measurements, a minimum of four different tumor areas were evaluated for each section, with a total of 400 (cell cycle) to 1000 (apoptosis) nuclei enumerated. All seminomas were positive for expression of p53 protein (>15% tumor cells positive). Tumor proliferation (MIB-1+) ranged from 2.3% to 22% in seminomas (mean 13.5%). Percent apoptotic nuclei ranged from 1.8 to 5.9% (mean 3.9%), with proliferation and apoptosis showing a positive correlation. All seminomas were negative for p21 expression, though normal testicular tissue had few strongly p21 + nuclei. PCNA expression was seen in 56-79% of tumor nuclei, indicating that replication complexes are likely not degraded in these tumors. These data indicate that seminomas have low to moderate proliferation and low apoptotic rates. Stabilization or overexpression of p53 and PCNA are seen in these tumors, and are unlikely related to cell cycle, or to p21 protein expression. Overall, these data indicate a significant perturbation of normal cell cycle regulatory pathways in these tumors.

Original languageEnglish (US)
Number of pages1
JournalBritish Journal of Urology
Volume80
Issue numberSUPPL. 2
StatePublished - Dec 1 1997
Externally publishedYes

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Germ Cell and Embryonal Neoplasms
Testis
Cell Cycle
Seminoma
Neoplasms
Proliferating Cell Nuclear Antigen
Apoptosis
Cell Cycle Proteins
Immunohistochemistry
Proteins
p53 Genes
Testicular Neoplasms
In Situ Nick-End Labeling
Paraffin
Anti-Idiotypic Antibodies
Cell Death
Antigens
Mutation

All Science Journal Classification (ASJC) codes

  • Urology

Cite this

Flanigan, R. C., Jin, J. Q., Challenger, B., Waters, W., Pyle, J. M., & Vincent, T. (1997). Cell cycle pathway perturbation in human germ cell tumors of the testis. British Journal of Urology, 80(SUPPL. 2).

Cell cycle pathway perturbation in human germ cell tumors of the testis. / Flanigan, Robert C.; Jin, Jm Quan; Challenger, Bob; Waters, W; Pyle, Joseph M.; Vincent, T.

In: British Journal of Urology, Vol. 80, No. SUPPL. 2, 01.12.1997.

Research output: Contribution to journalArticle

Flanigan, RC, Jin, JQ, Challenger, B, Waters, W, Pyle, JM & Vincent, T 1997, 'Cell cycle pathway perturbation in human germ cell tumors of the testis', British Journal of Urology, vol. 80, no. SUPPL. 2.
Flanigan RC, Jin JQ, Challenger B, Waters W, Pyle JM, Vincent T. Cell cycle pathway perturbation in human germ cell tumors of the testis. British Journal of Urology. 1997 Dec 1;80(SUPPL. 2).
Flanigan, Robert C. ; Jin, Jm Quan ; Challenger, Bob ; Waters, W ; Pyle, Joseph M. ; Vincent, T. / Cell cycle pathway perturbation in human germ cell tumors of the testis. In: British Journal of Urology. 1997 ; Vol. 80, No. SUPPL. 2.
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abstract = "Changes in expression of cell cycle-related proteins or their genes, and mechanisms regulating cell death (apoptosis) are now reported in many human solid tumors. Testicular germ cell tumors are unusual, in that most respond clinically to DNA damaging chemotherapeutic agents, suggesting that cell cycle and related apoptosis pathways are intact in these tumors. Since previous reports have indicated that mutations to the p53 gene are rare in these tumors, while expression of stable p53 protein is commonly seen, we have undertaken a study of cell cycle regulatory pathways to determine the defect or defects present. Paraffin-embedded tissue sections (5-6 micron) from 26 different testicular tumors have been evaluated to date, including 16 pure seminomas (stage I or II). Expression of cell cycle related proteins was determined by immunohistochemistry (IHC) using antigen retreival (TUF, PharMingen, 90° C for 30 min) and the following antibodies: anti-p53 (DO-7, DAKO), anti-PCNA (19A2, Coulter), Ki-67 (MIB-1, Immunotech), p21 (EA-10, CalBiochem). Apoptotic nuclei were evaluated using a modified TUNEL assay (Apo-BRDU, Phoenix Flow Systems). Tissues were lightly counterstained with hematoxalin. For all IHC measurements, a minimum of four different tumor areas were evaluated for each section, with a total of 400 (cell cycle) to 1000 (apoptosis) nuclei enumerated. All seminomas were positive for expression of p53 protein (>15{\%} tumor cells positive). Tumor proliferation (MIB-1+) ranged from 2.3{\%} to 22{\%} in seminomas (mean 13.5{\%}). Percent apoptotic nuclei ranged from 1.8 to 5.9{\%} (mean 3.9{\%}), with proliferation and apoptosis showing a positive correlation. All seminomas were negative for p21 expression, though normal testicular tissue had few strongly p21 + nuclei. PCNA expression was seen in 56-79{\%} of tumor nuclei, indicating that replication complexes are likely not degraded in these tumors. These data indicate that seminomas have low to moderate proliferation and low apoptotic rates. Stabilization or overexpression of p53 and PCNA are seen in these tumors, and are unlikely related to cell cycle, or to p21 protein expression. Overall, these data indicate a significant perturbation of normal cell cycle regulatory pathways in these tumors.",
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AU - Vincent, T.

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