P53-INDEPENDENT RESPONSE OF A HUMAN BREAST-CARCINOMA XENOGRAFT TO RADIOIMMUNOTHERAPY

BACKGROUND. Radiation-induced DNA damage resulting in p53 protein atta chment and downstream gene activation has been considered a major mech anism for tumor response to low dose rate radiation therapy. In this s tudy, the mechanism of tumor response, and p53 gene status as well as levels of expression of p53 pathway genes were investigated in a human breast tumor (HBT 3477) before and after yttrium-90-DOTA-peptide-ChL6 (Y-90-ChL6) treatment of these xenografts. METHODS. Mice with HBT 347 7 xenografts were treated with 260 mu Ci Y-90-ChL6 and sacrificed 3, 2 4 and 48 hours after injection. Reverse transcriptase-polymerase chain reaction and/or Western blotting were used to measure the tumor level s of p53, p21(CDKN1/WAF1) (p21), GADD45, and bcl-2. Single strand conf ormation polymorphism and direct sequencing were used to determine the mutational status of p53. Evidence of apoptosis was determined by cle avage of poly(ADP-ribose) polymerase (PARP). RESULTS. Tumors regressed 4-7 days after treatment with 260 mu Ci Y-90-ChL6, resulting in a 79% tumor response. The p53 gene mutation found at codon 342 in HBT 3477 resulted in truncation of the p53 protein, and correlated with undetec table basal p21 protein levels. GADD45 and p53 mRNA decreased after th erapy. bcl-2 mRNA was abundant, but decreased. Retinoblastoma phosphor ylation showed no changes. Cleavage of PARP was detected at 3 hours an d levels were increased greatly at 6 hours after therapy. CONCLUSIONS. Response in the Y-90-ChL6 treated HBT 3477 xenograft tumors was indep endent of p53 and occurred by apoptosis. The down-regulation of bcl-2 may be the key in this apoptotic response to low dose rate radioimmuno therapy. (C) 1997 American Cancer Society.