Induction and rejoining of DNA double-strand breaks in the lymphocytes of prostate cancer patients after radium-223 treatment as assessed by the γH2AX foci assay

Abstract

Aim The aim of this study is to assess if the number of radiation-induced double strand breaks (DSB) in lymphocytes of prostate cancer patients is affected after repeated Ra-223 therapies. In addition, we investigated the repair of ex vivo induced DSB to investigate the repair proficiency in patient’s lymphocytes over the therapy course.

Methods Before each of six therapy cycles, blood samples were obtained from seventeen patients. After separation of lymphocytes, the cells were subjected to immunofluorescence staining for detection of DSB-marking γH2AX foci. The number of foci per cell per patient sample was determined for each cycle (X1-X6, baseline foci per cell). Additionally, appropriate samples were exposed ex vivo to an X-ray dose of 1 Gy. The number of γH2AX foci per cell were analyzed after 0.5 h, 2 h and 24 h of recovery.

Results Patient-specific linear regression of the baseline foci per cell over the therapy cycles revealed no significant slopes in the regression lines. Likewise, the mean baseline foci per cell of all patients for cycles X2-X6 was not significantly elevated in comparison to the pre-therapeutic value (X1). The differences between the percentages of residual DSB and cycles were not significant, both at 2 h and 24 h repair time. Consideration of the X6/X1 ratios of both the number of lymphocytes and the amount of residual damage at 24 h indicated a significant correlation.

Conclusion Our findings indicate that the number of γH2AX foci per cell was not changed in dependence on the Ra-223 therapy cycles. The ability of patient’s lymphocytes to repair ex vivo induced DSB remained unaffected throughout the entire therapy course.

Zusammenfassung

Ziel Das Ziel dieser Studie ist die Analyse von strahleninduzierten Doppelstrangbrüchen (DSB) in Lymphozyten von Prostatakarzinompatienten im Verlauf der sechs Ra-223 Therapiezyklen (X1-X6). Die Reparaturkapazität der Lymphozyten wurde nach Applikation einer ex vivo Bestrahlung mit 1 Gy Röntgenstrahlung evaluiert.

Methoden Von 17 Patienten wurden vor jedem Therapiezyklus (X1-X6) Blutproben entnommen, die Lymphozyten isoliert und die DSB-assoziierten γH2AX Foci pro Zelle (Basiswerte) mit Immunfluoreszenzfärbung detektiert. Weiterhin wurden die entsprechenden Lymphozytenproben ex vivo mit 1 Gy Röntgenstrahlung bestrahlt und nach 0,5 h, 2 h und 24 h Reparaturzeit die γH2AX Foci pro Zelle bestimmt.

Ergebnisse Die Analyse der γH2AX Basiswerte für die einzelnen Patienten zeigte keine signifikanten Anstiege im Therapieverlauf. Die gemittelten γH2AX Basiswerte der Zyklen X2-X6 waren im Vergleich zum prätherapeutischen Wert (X1) nicht signifikant erhöht. Ein signifikanter Einfluss wiederholter Ra-223 Therapien auf die Reparatur ex vivo erzeugter DSB konnte nicht gefunden werden. Die Relation der Lymphozytenanzahl (X6/X1) korreliert mit der Relation residualer DSB (X6/X1) nach 24 h Reparaturzeit.

Schlussfolgerung Wiederholt applizierte Ra-223 Therapien zeigten mit dem verwendeten γH2AX Assay als DSB-Marker weder Auswirkungen auf die Basiswerte noch auf das Reparaturvermögen ex vivo induzierter DSB in Lymphozyten der Prostatakarzinom-Patienten.

Key words

prostate carcinoma - bone metastases - Ra-223 treatment - double strand breaks - γH2AX foci

Schlüsselwörter

Prostatakarzinom - Knochenmetastasen - Ra-223 Therapie - Doppelstrangbrüche - γH2AX Foci

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