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The causative factors leading to breast cancer are largely unknown. Increased incidence of breast cancer following diagnostic or therapeutic radiation suggests that radiation may contribute to mammary oncogenesis. This report describes the in vitro neoplastic transformation of a normal human mammary epithelial cell strain, 76N, by fractionated gamma-irradiation at a clinically used dose (30 Gy). The transformed cells (76R-30) were immortal, had reduced growth factor requirements, and produced tumors in nude mice. Remarkably, the 76R-30 cells completely lacked the p53 tumor suppressor protein. Loss of p53 was due to deletion of the gene on one allele and a 26-bp deletion within the third intron on the second allele which resulted in abnormal splicing out of either the third or fourth exon from the mRNA. PCR with a mutation-specific primer showed that intron 3 mutation was present in irradiated cells before selection for immortal phenotype. 76R-30 cells did not exhibit G1 arrest in response to radiation, indicating a loss of p53-mediated function. Expression of the wild-type p53 gene in 76R-30 cells led to their growth inhibition. Thus, loss of p53 protein appears to have contributed to neoplastic transformation of these cells. This unique model should facilitate analyses of molecular mechanisms of radiation-induced breast cancer and allow identification of p53-regulated cellular genes in breast cells.

BACKGROUND A prospectively applied treatment policy for breast‐conserving therapy used margin assessment as the exclusive guide to the intensity of radiation therapy directed at the tumor bed. METHODS From 1982 to 1994, 498 women with 509 Stage I/II breast carcinomas with a median follow‐up of 121 months were treated. Final margin status (FMS) categories were defined as greater than 5 mm, greater than 2–5 mm, greater than 0–2 mm, and positive. For margins less than or equal to 2 mm or indeterminate, reexcisions were performed if feasible. All patients received whole breast irradiation to 50.0–50.4 Gy. Final tumor bed boosts as a function of FMS were as follows: no residual on reexcision, no boost performed; FMS greater than 5 mm, boost of 10 Gy; FMS greater than 2–5 mm, boost of 14 Gy; FMS greater than 0–2 mm or positive, boost of 20 Gy. Cases were analyzed for local failure with respect to histology, age, tumor size, excision volume, reexcision, and total dose. RESULTS FMS was positive, greater than 0–2 mm, greater than 2–5 mm, and greater than 5 mm, and no residual tumor on reexcision in 21%, 20%, 17%, 14%, and 28% of cases, respectively. At 12 years, Kaplan–Meier local failure rates were 17% for FMS positive, 9% for FMS greater than 0–2 mm, 5% for FMS greater than 2–5 mm, 0% for FMS greater than 5 mm, and 6% for specimens without evidence of residuum on reexcision (P = 0.009). Patients 45 years old and younger had a 12‐year local failure rate of 15% whereas patients older than 45 years had a 12‐year local failure rate of 6% (P = 0.01). On multivariate analysis, young age (P = 0.03) predicted increased local failure rate, whereas margins that were less than or equal to 2 mm or positive predicted late (> 5 years) but not early (≤ 5 years) recurrence (P = 0.003). CONCLUSIONS Graded tumor bed dose escalation in response to FMS results in very low rates of local failure over the first 5 years for all FMS categories. However, tumors with close/positive margins have significantly increased local failure rates after 5 years of follow‐up even with increased radiation boost dose. In addition, graded tumor bed dose escalation does not fully overcome the adverse influence of young age. Cancer 2003;97:30–9. © 2003 American Cancer Society. DOI 10.1002/cncr.10981

The introduction and development of comprehensive two‐dimensional gas chromatography offers greatly enhanced resolution and identification of organic analytes in complex mixtures compared to any one‐dimensional separation technique. Initially promoted by the need to resolve highly complex petroleum samples, the technique’s enormous separation power and enhanced ability to gather information has rapidly attracted the attention of analysts from all scientific fields. In this Minireview, we highlight the fundamental theory, recent advances, and future trends in the instrumentation and application of comprehensive two‐dimensional column separation.