Abundance and subcellular localisation of cyclin D3 in human tumours

Abstract

The D-type cyclins are positive regulators of the GI phase of the mammalian cell cycle. Cyclins D I or D2 are over-expressed in several types of cancer, transform rodent cells in culture and therefore harbor hallmarks of cellular proto-oncogenes. In contrast, no data on expression of cyclin 0 3 in tissues and tumours are presently available. We have raised monoclonal antibodies (MAbs) specific for cyclin D3 and examined abundance and subcellular localisation of this GI cyclin in a series of human cultured cell types and in I80 primary tumours of diverse histogenesis. Cyclin D3 localised predominantly in nuclei of normal and tumour cells both in culture and in situ, and a pronounced cell-to-cell variation of its abundance was reminiscent of cyclins D I and D2. Immunohistochemical analysis of tumour and corresponding normal tissues showed strong aberrant accumulation of cyclin D3 in a subset (about 10%) of breast carcinomas, whereas only weak-to-moderate expression was found in colorectal, head and neck and uterine carcinomas, melanomas and soft tissue sarcomas. The specificity of the immunohistochemical data was confirmed by immunoblotting analysis of tissue and tumour lysates. Our results indicate that overabundance of cyclin D3 is considerably less frequent than that of cyclin DI, yet we identify subsets of breast tumours, and potentially lymphomas, as candidate tumour types with elevated cyclin D3 expression. o 1996 Wiley-Liss, Inc. Cyclins D1, D2 and D3 comprise a distinct subfamily of cyclin genes whose products positively regulate progression through the GI phase of the mammalian cell division cycle (reviewed by Sherr, 1994). The D-type cyclin proteins can combinatorially bind with several of the cyclin-dependent kinases (CDKs), in particular with CDK4 and CDK6 (re-viewed by Sherr, 1994; Hunter and Pines, 1994; Sherr and Roberts, 1995). According to current understanding, the cyclin D/CDK holo-enzymes control GI via phosphorylation of the retinoblastoma protein (pRB) (Kato et al., 1993; Lukas et al., 1995a; reviewed by Weinberg, 1995). This phosphorylation in mid-to-late GI cancels pRB's proliferation-constraining capacity through a conformational change resulting in release of a series of transcription factors from sequestration by the under-phosphorylated pRB. The liberated transcription factors , such as E2F, then regulate activities of a number of genes essential for initiation of DNA replication (reviewed by Weinberg, 1995). Expression of D-type cyclins is growth factor-inducible, thereby providing a link between various mitogenic signaling pathways and cell cycle machinery (Matsushime et al., 1991; Sherr, 1994). Consistent with the central role of the cyclin D family in growth control, and as a functional antagonist of the pRB tumour suppressor, a large body of evidence implicates deregulation of at least cyclin D1 as an oncogenic event involved in the pathogenesis of a wide range of neoplasms. Thus, the CCNDl gene on chromosome llq13, encoding cyclin D1, classifies as a proto-oncogene that can become...

Differential patterns of cell cycle regulatory proteins expression in transgenic models of thyroid tumours

“…Northern blot analysis of transcripts encoding the D-type cyclins showed that cyclin D3 transcripts were also the most abundant in the thyroids of control mice (Figure 3a). Immunohistochemistry (IHC) performed on thyroid tissue sections, revealed a nuclear localization of cyclin D3 but the staining appeared heterogeneous in intensity, as previously reported for cyclin D3 and other cyclin D subtypes in various normal and tumoral human tissues (Bartkova et al, 1996). About 23.3+3.5 (mean+s.d.…”

“…The antibodies used in this study were the mouse monoclonals DCS 6 (Lukas et al, 1994b), DCS 3.1 (Lukas et al, 1995b) and DCS 22 (Bartkova, 1996) respectively directed against human cyclin D1, D2 and D3; the mouse monoclonals DCS 31, DCS 32 and DCS 35 directed against human cdk4 (Bartek et al, manuscript in preparation); and the polyclonal rabbit serum against bovine thyroglobulin (Roger et al, 1985). Polyclonal rabbit sera raised against human cdk2 (M-2), rat cyclin E (M-20), mouse cyclin A (C-19) and human p27 kip1 (C-19), as well as the anti-human cdc2 mouse monoclonal (17) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).…”

Inhibition of cyclin D expression in human breast carcinoma cells by retinoids in vitro

“…Of these, increased expression of cyclins A, B, D and E have been reported in breast tumors or carcinoma cell lines (Buckley et al, 1993;Keyomarso and Pardee, 1993;Dutta et al, 1995;Keyomarsi et al, 1995). Within the cyclin D family, overexpression and/or ampli®cation of cyclin D1 (Gillett et al, 1994;Zhang et al, 1994;Zuckerberg et al, 1995;Courjal et al, 1996;Michalides et al, 1996) and D3 (Bartkova et al, 1996) have been reported in cohorts of invasive breast carcinomas; increased cyclin D1 expression was correlated with the presence of estrogen receptors. Both cyclin D1 transgenic mouse data (Wang et al, 1994) and transfection experiments using the human T-47D breast carcinoma cell line supported the hypothesis that cyclin D1 may be a breast cancer oncogene.…”

Expression of cyclin D1, D3, E, and p27 in human renal cell carcinoma analysed by tissue microarray

“…Cells with widespread alterations undergo apoptosis (Harbour and Dean, 2000). Different defects affecting the regulation of the G1/S transition have been detected in various malignancies (Bartkova et al, 1996;Gillet et al, 1996;Hommura et al, 2000;Kawauchi et al, 2001). This suggests that cell cycle aberrations probably represent one of a limited number of key events in the transformation process (Hahn et al, 1999).…”