Guoren Deng

Loss of Heterozygosity in Normal Tissue Adjacent to Breast Carcinomas

Loss of heterozygosity (LOH) was detected in morphologically normal lobules adjacent to breast cancers. The most frequent aberration was at chromosome 3p22-25; of ten cases with this LOH in the carcinoma, six displayed the same LOH in adjacent normal lobules. This suggests that in a subset of sporadic breast cancers, a tumor suppresser gene at 3p22-25 may be important in initiation or early progression of tumorigenesis. Among sixteen breast cancers with LOH at 17p13.1 and five breast cancers with LOH at 11p15.5, one case each displayed the same LOH in adjacent normal lobules. Thus the molecular heterogeneity that characterizes invasive breast cancers may occur at the earliest detectable stages of progression.

Fine needle aspiration techniques for the characterization of breast cancers.

Fine needle aspiration biopsy (FNAB) is a well established method for diagnosing breast lesions, including cancers. FNAB does not require surgery and uses only a small amount of material. FNAB can also be used to acquire material for special studies. This is especially useful with small tumors (≦ 1 cm) when most of the material is needed to make a histologic diagnosis. Immunostaining techniques can be used on FNABs to investigate proliferation by bromodeoxyuridine uptake or Ki‐67 labeling. Immunostaining techniques can also be used to identify oncoprotein expression, such as of p53. Fluorescence in situ hybridization is a technique that can be used to gather cytogenetic information directly from interphase tumor cells and is well suited for use with FNAB material because the harvested nuclei are intact and no cumbersome dissociation processing is needed. Flow cytometric techniques can be applied to FNAB material to study DNA content and S‐phase fraction. Material acquired by FNAB can also be analyzed by the polymerase chain reaction followed by mutation detection. In this report, the authors show the applicability of these various analytic approaches to FNAB material from primary breast cancers. They show that it is essential that the FNAB harvest is representative, ample, and well prepared for the success of these studies.

Amplifications of oncogeneerbB-2 and chromosome 20q in breast cancer determined by differentially competitive polymerase chain reaction

SummaryA new method of measuring gene copy number in small samples of DNA was used to measure amplification of theerbB-2 gene and of chromosome 20q in breast cancers. This method, termed ‘differentially competitive polymerase chain reaction’ (DC-PCR) combines the advantages of two other techniques for measuring amplification by PCR, namely differential PCR and competitive PCR. The DC-PCR methodology was evaluated for sensitivity and specificity by comparing amplification oferbB-2 measured by DC-PCR with that obtained by fluorescencein situ hybridization (FISH) for 42 cases or Southern blotting and/or slot blot analysis for 34 cases. There was over 90 percent concordance with both FISH and Southern blotting and/or slot blot analysis.DC-PCR was used to further characterize the newly described amplicon at chromosome 20q. By analyzing DNA from 10 breast cancer cell lines at 7 different loci, we identified a potential common region of amplification of approximately 5 centimorgans at chromosome 20q13 bordered by loci D20S52 and RMC20C001-S1. One hundred and seventeen cases of primary breast cancer were evaluated for amplification at these two loci. Amplification at one or more loci, defined as > 1.5 fold higher copy number than that of normal DNA, was found in 25 cases (21%). Sixteen cases were amplified at only one of the two probes (12 cases for RMC20C001-S1 and 4 cases for D20S52), suggesting that the target gene lies between the two markers or that there are two independent target genes within a small chromosome region.