Summary
Purpose
Inflammatory breast cancer (IBC) is the most aggressive form of locally advanced breast cancer (LABC). The IBC phenotype is characterized by an infiltrative growth pattern, increased (lymph)angiogenesis and the propensity to invade dermal lymphatics. In pancreatic cancer, interactions between caveolin-1 and RhoC GTPase, a key molecule in causing the IBC phenotype, regulate tumour cell motility and invasion. In this study we sought to investigate the role of caveolin-1 and -2 in IBC cell lines and in human IBC samples.
Experimental design
Differential methylation techniques identified the methylation status of the caveolin-1 and -2 promoters in human mammary epithelial cells (HMECs) and the SUM149 cell line. In cell line experiments, caveolin-1 and -2 mRNA and protein expression were compared in HMECs, MCF10A, the SUM102 non-IBC cell lines and 2 IBC cell lines (SUM149 and SUM190). Furthermore, caveolin-1 and -2 mRNA and protein expression were compared in human IBC and non-IBC samples using cDNA microarray, real-time qRT-PCR and immunohistochemistry. Results were correlated with RhoC protein expression data.
Results
In the SUM149 cell line, the caveolin-1 and -2 promoter sites were hypomethylated. A significantly increased expression of caveolin-1 and -2, both at the mRNA and protein level was found in IBC cell lines and in human samples of IBC: caveolin-1 and -2 mRNA were respectively 1.7 (p = 0.02) and 2.2 (p = 0.03) fold more expressed in IBC compared to non IBC and at the protein level, 41.4% of IBC specimens expressed either caveolin-1 or -2, compared to 15.6% of non-IBC specimens (p = 0.03). Furthermore a correlation was found between RhoC protein expression and caveolin-1 (p = 0.1) or caveolin-2 (p = 0.09) or either caveolin-1 or -2 protein expression (p = 0.04).
Conclusions
Although considered a tumour suppressor in breast cancer, we demonstrated overexpression of caveolin-1 and -2 in IBC cell lines and in human samples of IBC, most likely due to hypomethylation of their respective promoters. These results confirm the distinct molecular signature of IBC. Our data further suggest interaction between RhoC GTPase and the caveolins in IBC.
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Acknowledgments
G. Van den Eynden is a research assistant of the Fund for Scientific Research Flanders. This work was supported by FWO Flanders ‘Kom op tegen Kanker’, Grant No. G. 0330.02. and FWO ‘Kredieten aan Navorsers’ Grant No. 1.5.158.03. We thank AL Harris, M.D. PhD and SB Fox, M.D. PhD for the cooperation and support in the cDNA microarray experiments and we thank Min Lin, M.D. and H. Elst, LT for expert technical assistance. Furthermore we want to thank the technical staff of the laboratories of pathology from the University Hospital Antwerp and the General Hospital Sint-Augustinus.
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Address for offprints and correspondence: Peter B. Vermeulen, Department of Pathology, AZ St.-Augustinus, Oosterveldlaan 24, B-2610 Wilrijk, Belgium; Tel.: +32-3-443-4608; Fax: +32-3-443-3036; E-mail: peter.vermeulen@gvagroup.be
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Van den Eynden, G.G., Van Laere, S.J., Van der Auwera, I. et al. Overexpression of caveolin-1 and -2 in cell lines and in human samples of inflammatory breast cancer. Breast Cancer Res Treat 95, 219–228 (2006). https://doi.org/10.1007/s10549-005-9002-1
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DOI: https://doi.org/10.1007/s10549-005-9002-1