Overexpression of cyclins D1 and D3 during estrogen-induced breast oncogenesis in female ACI rats

Open Access
Publisher Website
Google Scholar
Abstract
A common feature of human breast oncogenesis is cell cycle deregulation. The expression of cyclins D1 and D3 was examined during estradiol-17β (E 2 )-induced mammary tumorigenesis in female August Copenhagen Irish (ACI) rats. Low serum E 2 levels (∼60–120 pg/ml) were sufficient to induce mammary gland tumors (MGTs) that remarkably resemble human ductal breast cancer (BC) at the histopathologic and molecular levels. Western blot analysis of the E 2 -induced MGTs revealed a marked rise in cyclins D1 (24-fold), D3 (9-fold) and cdk4 (3-fold) expression compared with age-matched untreated controls. Small focal dysplasias with large, pale staining nuclei were commonly seen at 3–3.6 months, large focal dysplasias, including atypical ductal hyperplasia at 3.6–4.3 months, ductal carcinoma in-situ (DCISs) at 4.3–5.0 months, and 100% incidence of invasive ductal BC/frank tumors at 5–6 months were detected after E 2 treatment. Immunohistochemical analysis of serial sections of focal dysplasias, DCISs and invasive ductal carcinomas showed overexpression of cyclins D1, D3, estrogen receptor-α (ERα) and progesterone receptor (PR). However, cyclin D3 expression, unlike D1, was confined essentially to early pre-malignant lesions (focal dysplasias and DCISs) and primary MGTs with 2 -induced MGTs and their binding to cdk4 was significantly elevated. Semi-quantitative reverse transcriptase PCR analysis of the E 2 -induced MGTs exhibited increased expression of cyclins D1 (2.9-fold) and D3 (1.4-fold) mRNA, indicating that their elevated protein expression was due in part to an increase in mRNA transcription. However, when analyzed by quantitative real-time Q-PCR, these genes were not amplified. These data indicate that in female ACI rat mammary glands, E 2 -induced pre-malignant lesions differentially and selectively express cyclins D1 and D3, thus contributing to a distinct growth advantage of these pre-neoplasias relative to E 2 -elicited normal hyperplasia.