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Controversy exists regarding the relationship of the degree of c-erbB-2 amplification to other prognostic factors in breast cancer. To determine the degree of amplification of c-erbB-2 exactly, a sensitive and quantitative method is required. We have developed a competitive PCR method to quantitatively determine the amplification of the c-erbB-2 oncogene. Using this method, we evaluated DNA from 27 breast cancer tissue specimens and DNA from peripheral blood leukocytes from a normal individual. Regarding the relationship between the degree of c-erbB-2 amplification and clinicopathological factors, we found a greater degree of amplification of the c-erbB-2 oncogene in estrogen receptor- negative or progesterone receptor-negative specimens than in positive ones and in lymph node metastasis-positive specimens than in negative specimens, in stages II, III, and IV of disease compared with stage I disease, and in samples with positive lymphatic vessel invasion than with no lymphatic vessel invasion. Generally, these factors were seen in the group of patients who had a bad prognosis. By univariate analysis and multivariate analysis, reverse correlation was observed between amplification of c-erbB-2 and overall survival. Regarding disease-free survival, these relationships were observed only with univariate analysis in our group of patients.

The formation of microvessels in tumors by angiogenesis is considered to be an important prognostic factor, and closely correlates with lymph node metastasis. We used color Doppler ultrasound to examine the relationship between the amount of blood vessels in tumors and pulsatility index (PI), and tumor size in breast cancers, with and without regional lymph node metastasis. Doppler ultrasound was performed on 80 patients with breast cancer prior to surgery. The concentration of vascular endothelial growth factor (VEGF) within the tumors was measured following surgery in 42 cases chosen at random. In the negative metastatic nodes group, the number of vessels in the tumor correlated positively with tumor diameter. In the positive metastatic nodes group, however, the number of blood vessels in the tumor did not correlate with tumor diameter. Differences in tumor vascularity between node positive and negative groups were useful in determining the status of node metastasis in subsequent analysis. Fifteen of 17 cases of tumors that measured20 mm, and in which there were no blood vessels, were node-negative. There were no node-negative tumors measuring20 mm in diameter (p=0.003). Conversely, in nodes with positive metastasis, blood vessels were observed in 5 of 7 tumors that measured15 mm in diameter (p=0.019). These findings may be useful in estimating the likelihood of metastasis to regional lymph nodes. PI was directly proportional to tumor size in the negative nodes group (r=0.47). There was no such correlation in the positive nodes group. There was no correlation between VEGF concentration in the tumor and the number of blood vessels in that tumor. In conclusion color Doppler analysis of blood vessels appears to be useful in predicting lymph node metastasis, especially for small tumors.

A 74-year-old woman complained of a small nodule in the outer lower quadrant of her left breast. On physical examination, a 0.9 x 0.8 cm, round-shaped and firmly elastic nodule was palpated. Excisional biopsy was performed. Histologically, the tumor was separated into, with a bicellular pattern, containing both numerous glandular structures and numerous spindle-shaped cells. Immunohistochemical staining for EMA and cytokeratin showed strongly positive immunoreactivity for epithelial cells. Staining foraipha;-SMA showed strongly positive immunoreactivity for myoepithelial components. Staining for keratin and S-100 protein showed weakly positive immunoreactivity for myoepithelial cells. Microscopically, the tumor was diagnosed as adenomyoepithelioma of the breast. Immunohistochemical examination is needed to distinguish epithelial cell proliferation from myoepithelial cell proliferation. Immunohistochemical examination using antibodies against EMA, alpha-SMA, Keratin, cytokeratin and S-100 protein, is indispensable.