A Tissue Graft Model of DNA Damage Response in the Normal and Malignant Human Prostate

Abstract

Purpose DNA damage responses are relevant to prostate cancer initiation, progression and treatment. Few models of the normal and malignant human prostate that maintain stromal-epithelial interactions in vivo exist in which to study DNA damage responses. We evaluated the feasibility of maintaining tissue slice grafts at subcutaneous vs subrenal capsular sites in RAG2−/− γC−/− mice to study the DNA damage responses of normal and malignant glands. Materials and Methods We compared the take rate and histology of tissue slice grafts from fresh, precision cut surgical specimens that were maintained for 1 to 4 weeks in subcutaneous vs subrenal capsular sites. Induction of γH2AX, p53, ATM and apoptosis was evaluated as a measure of the DNA damage response after irradiation. Results The take rate of subcutaneous tissue slice grafts was higher than typically reported but lower than at the subrenal capsular site. Subcutaneous tissue slice grafts frequently showed basal cell hyperplasia, squamous meta-plasia and cystic atrophy, and cancer did not survive. In contrast, normal and malignant histology was well maintained in subrenal capsular tissue slice grafts. Regardless of implantation site the induction of γH2AX and ATM occurred in tissue slice graft epithelium 1 hour after irradiation and decreased to basal level by 24 hours, indicating DNA damage recognition and repair. As observed previously in prostatic ex vivo models, p53 was not activated. Notably, tumor but not normal cells responded to irradiation by undergoing apoptosis. Conclusions To our knowledge this is the first study of DNA damage responses in a patient derived prostate tissue graft model. The subrenal capsular site of RAG2−/− γC −/− mice optimally maintains normal and malignant histology and function, permitting novel studies of DNA damage responses in a physiological context.

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“…In addition, we selected tumor cell lines due to the ease of calculating SF in proliferating cells. Therefore, our results may not necessarily apply to peripheral blood lymphocytes or normal human cells, although there appear no data suggesting quite different patterns in DSB repair between normal cells and tumor cells [ 18 , 39 , 47 ]. Ideally, similar experiments should also be performed in peripheral blood lymphocytes and normal human cells to assess the adverse effects of CT examinations.…”

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