Intratumoral heterogeneity was studied in human breast cancer by exami
ning separate tumor lesions of individual patients. Tumor samples were
obtained from each patient by fine-needle biopsies from 2 to 4 separa
te tumor lesions. We used a semi-quantitative PCR to distinguish betwe
en samples with gene amplification and single gene copy samples. Five
genes were analyzed in each biopsy: MDR-1, dihydrofolate reductase, th
ymidylate synthase, c-erb-B2 and int-2. Three groups of patients were
examined: those awaiting initiation of treatment; those receiving firs
t-line endocrine therapy; and those receiving second-line endocrine tr
eatment. A pattern of intratumoral heterogeneity for gene amplificatio
n was clearly apparent. The frequency of amplification was lowest befo
re initiating therapy and highest in patients receiving second-line tr
eatment (p = 0.023). (C) 1994 Wiley-Liss, Inc.