In vivo tumor measurement of DNA damage, DNA repair and NAD pools as indicators of radiosensitization by metoclopramide.
Metoclopramide (MCA), a N-substituted benzamide, causes DNA strand breaks and inhibits DNA repair in vitro and sensitizes radiation and chemotherapeutic drugs in human squamous cell carcinomas when xenographed into nude mice or in a rat glioma model. Here we report on the evaluation of the mechanism behind the radiosensitizing effects of MCA. DNA damage was measured in vivo in a CBA-mouse tumor line (A12B3, sarcoma tumor) by using both alkaline elution and nucleoid sedimentation analysis of cell suspensions prepared from either resected tumor, spleen tissues or whole blood samples. The amount of DNA damage caused by radiation alone, measured 30 min after the irradiation was started, was dose dependent up to 18 Gy in all tissues. The radiation-induced DNA damage in tumor tissue was elevated compared to radiation alone in the presence of MCA, but the level was not higher at 18 Gy compared to 6 Gy in the presence of MCA, and it was still not fully repaired 12 h after irradiation. HPLC analysis of the NAD pools in tumor tissue after DNA damage induction showed a delay in the recovery of the NAD pools (presumably due to the presence of still unrepaired DNA) after exposure to MCA (2 mg/kg) + radiation (6 Gy) compared to tumors exposed to radiation (6 Gy) only, which were fully restored after 48 h. These data confirm earlier published in vitro data on MCA as an inducer of DNA damage and an effector of DNA repair. In addition, the in vivo measurement of radiation-induced DNA damage and DNA repair using the nucleoid sedimentation and alkaline elution assays together with NAD pool determinations may prove to be effective intermediate endpoints in the evaluation of drugs as potential radiosensitizers.
Olsson
A
A
Department of Molecular Ecogenetics, University of Lund, Sweden.
Sheng
Y
Y
Kjellén
E
E
Pero
R W
RW
D000818:Animals; D002588:Cesium Radioisotopes; D004247:DNA / Q000187:drug effects* / Q000528:radiation effects; D004249:DNA Damage; D004260:DNA Repair / Q000187:drug effects* / Q000528:radiation effects; D004273:DNA, Neoplasm / Q000187:drug effects* / Q000528:radiation effects; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D006801:Humans; D007700:Kinetics; D008787:Metoclopramide / Q000494:pharmacology*; D051379:Mice; D008808:Mice, Inbred CBA; D009243:NAD / Q000032:analysis / Q000378:metabolism*; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*; D051381:Rats; D011923:Rats, Nude; D012513:Sarcoma, Experimental / Q000235:genetics / Q000378:metabolism*; D013154:Spleen / Q000187:drug effects* / Q000378:metabolism / Q000528:radiation effects; D013997:Time Factors
D002588:Cesium Radioisotopes; D004273:DNA, Neoplasm; D011838:Radiation-Sensitizing Agents; D009243:NAD; D004247:DNA; D008787:Metoclopramide