title: Oncogenes and cellular-sensitivity to radiotherapy - a study on murine keratinocytes transformed by v-h-ras, v-myc, v-Neu, adenovirus e1a and mutant p53.
abstract: Mechanisms involved in cellular radioresistance are mostly unknown and may be related to specific genetic alterations. In order to correlate the most frequent oncogenic alterations detected in tumors and ionizing radiation resistance, we studied the effect of irradiation on murine keratinocytes transformed by different oncogenes. Mouse PAM 212 keratinocytes were transformed by transfection or retroviral mediated infection with the oncogenes v-H-ras, v-myc, adenovirus Ela, neu and a mutant p53 (mp53). Cells were gamma irradiated with a Co-60 source. Cell viability was evaluated by the crystal violet method and thymidine uptake and data adjusted to the linear-quadratic model. Surviving fraction 2Gy (SF2) and DO was calculated. Cell cycle study was assessed by incorporation of bromodeoxyridine (BrdUrd) and flow cytometry. p53 protein was studied by Western-blot and apoptosis in DNA agarose gels. The surviving fraction for the different keratinocytes, PAM 212, 212 neo, 212 Ela, 212 v-H-ras, 212 myc, 212 neu and 212 mp53 was 0.79, 0.78, 0.34, 0.82, 0.68, 0.74, and 0.72, respectively. Ela oncogene induced a great sensitivity to irradiation and v-H-ras a mild radioresistance. In flow cytometry, 212 Ela keratinocytes displayed a pronounced and prolonged arrest in G2/M phase. Apoptosis was observed after irradiation only in the 212 Ela keratinocytes. With these results, we conclude that some oncogene products may modulate radiosensitivity in keratinocytes. Mechanisms involved in radiosensitivity mediated by the Ela oncogene seem to be related to p53 protein level, induction of apoptosis and to an irreversible premitotic arrest in G2/M phase, ineffective for repair of DNA damage.
authors.affiliation: CLIN PUERTA DE HIERRO,DEPT PATHOL,E-28035 MADRID,SPAIN. INST INVEST BIOMED,E-28035 MADRID,SPAIN.
authors.forename: E
authors.initials: E
authors.lastname: Marchetti
authors.forename: J
authors.initials: J
authors.lastname: Romero
authors.forename: R
authors.initials: R
authors.lastname: Sanchez
authors.forename: J
authors.initials: J
authors.lastname: Vargas
authors.forename: C
authors.initials: C
authors.lastname: Dominguez
authors.forename: J
authors.initials: J
authors.lastname: Lacal
authors.forename: S
authors.initials: S
authors.lastname: Ramon y Cajal