Epstein-Barr virus gene expression in human breast cancer: protagonist or passenger?

The presence and transcriptional expression of Epstein-Barr virus (EBV)-encoded genes, oestrogen receptor (ER) status and degree of lymphocyte infiltration were evaluated in 15 mastectomy-removed breast cancer samples, mostly of ductal origin. With regard to these parameters, the tumours were heterogeneous. Viral genes, including EBNA1 - a universal EBV marker - and others, selected in part on the basis of expression in other EBV-associated carcinomas and/or presence in an epithelial cell immortalising subfragment p31 of viral DNA, were detected in up to 40% of the breast malignancies. The small viral RNAs, EBERs, were not observed. In culture, p31 EBV DNA, alone among EBV fragments, stimulated the growth of human breast-milk epithelial cells. There was no correlation between viral and ER expression and tumours were heterogeneous with regard to their invasive lymphocytes: of three studied in detail, one contained none, another had (mainly) T-lymphocyte aggregates on the tumour periphery, and a third (BC 12) was infiltrated with both T- and B-lymphocytes. BC 12 differed in several aspects from other malignancies in expressing a transcriptional activator (BZLF1) associated with overcoming virus latency, and failing to express a viral oncogene, BARF1. Arguments are given for EBV as a protagonist cocarcinogen in some breast malignancies.

A comparison of immunohistochemical markers of cell proliferation with experimentally determined growth fraction.

The relationship between immunoreactivity for cell proliferation markers (Ki67 and PCNA) and the growth fraction as determined by the fraction of labelled mitoses method was assessed in xenograft tumours grown from the LoVo cell line in nude mice. FLM curves were constructed by injecting tritiated thymidine and then preparing autoradiographs from the tumours. From this data an estimate of growth fraction and cell cycle time were made. Using frozen material from the same tumours, the Ki67 index was determined by immunostaining. PCNA staining was determined in the fixed material which had been used for the autoradiographs. The results show that Ki67 staining follows the same trend as the FLM-determined growth fraction as the tumour increases in size and the rate of growth decreases. However the Ki67 index does produce a consistent overestimate of the growth fraction in this in-vivo system, as compared to observations in cell culture. PCNA staining showed virtually 100 per cent positive staining in all the tumours, which is likely to reflect the long half-life of the antigen, compared to the very fast cell-cycle time of the xenograft tumours. These results show that staining with proliferation markers is not a precise determinant of growth fraction. Ki67 staining is a method that can be usefully applied as an operational marker of cell proliferation, but should not be used uncritically. Further caution is necessary in the use of PCNA. The findings also demonstrate the need to use a range of methods when assessing a new proliferation marker.

c-erbB-2 oncogene expression in ovarian cancer.

One hundred and four common epithelial malignancies of the ovary were stained for c-erbB-2 using an affinity-purified polyclonal antibody 21N. Twenty-three out of 104 (22.1 per cent) showed cytoplasmic staining alone. Nine out of 104 (8.7 per cent) showed both membrane and cytoplasmic staining. In a multivariate Cox analysis with other known risk factors the relative risk for cytoplasmic staining alone was 1.456 (chi 2 = 1.71, P greater than 0.1) and for membrane and cytoplasmic staining 0.316 (chi 2 = 7.95, P less than 0.005). These results do not support an adverse prognostic effect of c-erbB-2 in our patients.