Topoisomerase II expression as a determinant of chromosomal radiosensitivity and possible susceptibility in breast cancer
© BioMed Central Ltd 2008
Published: 13 May 2008
Elevated chromosomal radiosensitivity in lymphocytes of breast cancer patients is thought to be an indicator for the presence of one or more as yet unidentified genes of low penetrance that promote susceptibility to the disease in up to 60% of cases [1, 2]. One such gene may be TOPO2A, encoding for the DNA processing enzyme topoisomerase IIa. The involvement of topoisomerase IIa is predicted from the author's model for formation of chromatid breaks . In the model the DNA double-strand break is not directly involved in the chromatid break, but acts as an initiator in a sequence of events leading to a chromatid break. It is thought that a chromatid break may be formed by a misjoining of chromatin ends during topoisomerase IIa decatenation of chromatids as the cell progresses through G2 towards mitosis. Topoisomerase IIa is known to be vulnerable to perturbation by oxidative stress during the precise process of cutting and joining DNA strands .
Gamma-radiation-induced chromatid breaks are scored for chromatid breaks in colcemid-blocked chromosome spreads of metaphase HL60 and mitoxantrone-resistant variants: MX1 and MX2 cells with reduced topoisomerase II expression. Topoisomerase IIa expression levels were measured using western blotting. SiRNA was used to knock down expression in normal exponentially growing human cells that are irradiated with a low dose of γ-rays and scored for the presence of chromatid breaks. The chromatid break frequency and topoisomerase IIa expression (ELISA assay) are being compared in 3-day-stimulated peripheral blood T lymphocytes from a group of breast cancer patients and control individuals exposed to a low dose of γ-radiation.
We show that chromatid radiosensitivity (based on the frequency of metaphase chromatid breaks in irradiated G2 cells) is significantly lower in a topoisomerase IIa underexpressing variant cell lines , and preliminary results show that reducing expression with SiRNA also reduces chromatid radiosensitivity. In a pilot study we are currently comparing the chromatid radiosensitivity and topoisomerase IIa expression in stimulated peripheral blood lymphocytes of a group of Tayside breast cancer patients and a similar number of normal noncancer control individuals.
Our data support the hypothesis that topoisomerase IIa expression is a determinant of chromatid break frequency in irradiated G2 cells, and thus could be an underlying cause of the observed variability of chromatid radiosensitivity among both sporadic breast cancer cases and normal control individuals.
Supported by the CSO and Breast Cancer Campaign.
- Scott D, Barber JBP, Spreadborough AR, Burrill W, Roberts SA: Increased radiosensitivity in breast cancer patients: a comparison of two assays. Int J Radiat Biol. 1999, 75: 1-10. 10.1080/095530099140744.View ArticlePubMedGoogle Scholar
- Riches AC, Bryant PE, Steel CM, Gleig A, Robertson AJ, Preece PE, Thompson AM: Chromosomal radiosensitivity in G2-phase lymphocytes identifies breast cancer patients with distinctive tumour characteristics. Br J Cancer. 2001, 85: 1157-1161. 10.1054/bjoc.2001.2086.View ArticlePubMedPubMed CentralGoogle Scholar
- Bryant PE: Repair and chromosomal damage. Radiother Oncol. 2004, 72: 251-256. 10.1016/j.radonc.2004.07.005.View ArticlePubMedGoogle Scholar
- Chen LT-K, Yu C, Mao Y, Wang H, Liu LF: Activation of topoisomerase II mediated excision of chromosomal DNA loops during oxidative stress. Genes Dev. 1999, 13: 1553-1560. 10.1101/gad.13.12.1553.View ArticlePubMedPubMed CentralGoogle Scholar
- Terry S, Riches AC, Bryant PE: A role for topoisomerase IIa in the formation of radiation-induced chromatid breaks. Br J Cancer. 2008.Google Scholar