Claudio Dati

Tamoxifen up-regulates c-erbB-2 expression in oestrogen-responsive breast cancer cells in vitro

Expression of the c-erbB-2 proto-oncogene is inhibited by oestrogens in oestrogen-responsive human breast cancer cells, at both mRNA and protein level. Here we report that, where the regulation of c-erbB-2 is concerned, tamoxifen displays a full anti-oestrogenic activity, enhancing the expression of c-erbB-2 in oestrogen receptor-positive cells cultured with untreated fetal calf serum or reversing the inhibitory effect of added oestrogens. Meanwhile, tamoxifen strongly inhibited cell growth. Tamoxifen was inactive on both c-erbB-2 expression and growth of oestrogen receptor-negative cells. These results may have important implications to explain occasional failure of tamoxifen therapy in oestrogen receptor-positive breast cancers.

Hormonal regulation of c-erbB-2 oncogene expression in breast cancer cells

Expression of the c-erbB-2 (neu, HER-2) oncogene is found to be subjected to hormonal and developmental regulation in normal as well as neoplastic mammary cells. We have previously reported that estrogens inhibit c-erbB-2 expression at both the mRNA and protein level in estrogen receptor (ER)-positive, but not in ER-negative, breast cancer cell lines. Reversion of c-erbB-2 inhibition is seen with tamoxifen. The effect on c-erbB-2 expression of several other hormones and factors, which influence mammary cell growth and differentiation, has been studied. Our observations indicate that, in normal and neoplastic mammary cells, c-erbB-2 expression is inversely related to cell proliferation. While estrogens, anti-estrogens and cAMP clearly regulate c-erbB-2 mRNA levels, epidermal growth factor dramatically decreases the c-erbB-2 protein without affecting the level of c-erbB-2 mRNA. Therefore, different signals converging in terms of cell proliferation regulate c-erbB-2 expression by different molecular mechanisms.

ErbB‐4 and neuregulin expression in the adult mouse olfactory bulb after peripheral denervation

ErbB‐4 is expressed by the periglomerular and the mitral/tufted cells of the adult mouse olfactory bulb (OB) and in the present work we tested whether this expression is regulated by the olfactory nerve input to the OB. Reversible zinc sulphate lesions of the olfactory mucosa were made in adult mice and the deafferented OB analysed by immunohistochemistry, Western blotting and semiquantitative RT‐PCR. Following deafferentation, the expression of erbB‐4, erbB‐2 and neuregulin‐1 (NRG‐1) mRNAs in the OB was altered. At early stages (7–14 days) after lesion the levels of expression of olfactory marker protein (OMP), tyrosine hydroxylase (TH), erbB‐4 and NRG‐1 mRNAs were decreased, whilst expression of erbB‐2 increased and that of NRG‐2 was not significantly altered. We observed at least two distinct time courses for these expression changes. The lowest amounts of mRNA for erbB‐4 and NRG‐1 were observed at day 7 after lesion, whilst mRNAs for TH and OMP were lowest at day 14. At day 28 after the lesion, when olfactory receptor neuron axons had reinnervated the olfactory bulb, the expression levels of OMP, TH, erbB‐2, erbB‐4 and NRG‐1 were identical to control values. These results indicate that the expression of erbB‐4 mRNA and protein in periglomerular and mitral cells is controlled by peripheral olfactory innervation. The tight correlation in NRG‐1 and erbB‐4 expression levels also suggests a possible functional link that deserves further exploration.

Hormonal Regulation of Type I Receptor Tyrosine Kinase Expression in the Mammary Gland

Hormones guide mammary gland development and differentiation by regulating the expression of local growth factors and their receptors at the cell surface. In line with this principle the expression of the epidermal growth factor receptor (EGFR)3 and ErbB2 receptors varies in the mammary gland during pregnancy, following the changing hormonal profile. In breast cancer, expression of EGFR and ErbB2 is clearly related to the absence of estrogen and progesterone receptors. In breast cancer cells in vitro, the expression of these receptors is modulated by hormones and other growth-modulatory reagents. Moreover, transcriptional regulation of both EGFR and ERBB2 by estrogens has been demonstrated. The action of hormones may therefore result in the differential availability of individual ErbB family members at the cell surface, in this way determining the specific response of the cell to EGF-like factors and heregulins.

Hormonal control of growth factor receptor expression

In this report, we have discussed a series of results obtained in our laboratory that, together with data by other authors, demonstrate that the expression of the erbB-2 tyrosine kinase receptor oncogene in breast cancer cells is regulated by multiple factors and hormones, which modulate their growth and differentiation. In particular, we have shown that estrogens specifically inhibit erbB-2 expression by transcriptional repression, which is exerted through a sequence within the erbB-2 gene promoter. Estrogens control mammary cell growth directly, by inducing early gene expression, and indirectly, by increasing autocrine growth factor production or decreasing growth inhibitors. The data presented here suggest that mammary cells respond to estrogen also by modifying the receptor array on their surface, thus setting their own sensitivity to the different autocrine and paracrine factors. As a first consequence, the modulation of erbB-2 expression level by antiestrogen may represent a point to consider when selecting breast cancer patients for hormonal therapy, in those (few) cases where estrogen receptor positivity accompanies erbB-2 amplification. On the other hand, antiestrogen-induced upregulation of erbB-2 may improve tumor targeting of drugs designed to interact or interfere with erbB-2, such as humanized antibodies, immunotoxins, or engineered ligands. These possibilities should be tested in appropriate model systems in the future.