Article
Genetic profiling of breast cancer brain metastases: The MDA-MB 231 array experiment
Search Medline for
Authors
Published: | September 16, 2010 |
---|
Outline
Text
Objective: Breast cancer is the second most common cause of brain metastases. We report on the genetic profiling of brain- and bone-selective invasive and metastatic MDA-MB-231 breast cancer cells using cDNA Array technology, real-time RT-PCR and protein detecting methods.
Methods: Gene profiling was performed using the 16K gene chip of the National Human Genome Research Institute, NIH, USA. Results were verified by real-time RT-PCR. Protein expression was examined using enzyme-linked immunosorbent assays (ELISA), western blotting and immunohistochemistry. Migration and invasion assays were performed with the QCM 96-well Migration/Invasion Assay +/- specific inhibitors.
Results: cDNA array analysis revealed significant over- or underexpression in brain- and / or bone-seeking cells of 113 genes. 9 gene sequences were chosen for data validation using real-time RT-PCR. Significant changes in both selective clones were found for: (1) matrix-metallo-proteinase 1 (MMP-1). Further examination revealed increased expression of MMP-1 and -9 in both selective clones on the mRNA and protein level. (2) metastasis suppressor gene KISS-1, Further examination of human tissue samples revealed reduced expression of several metastasis suppressor genes in breast cancer brain metastases versus primary tumors and normal breast tissue: Kai1, BRMS-1, Mkk-4, Maspin (SERPIN B5). (3) vascular factors endoglin and TIE-1. Endoglin was highly overexpressed in both selective clones and, moreover, was associated with matrix-metallo-proteinases -1 and -19 and was also associated with the cells migration and invasion capacity.
Conclusions: Several genes show significant over- or underexpression in brain-selective metastatic MDA-MB-231 clones. Herein, metastasis suppressor genes KiSS1, Kai1, BRMS-1, Mkk-4 and Maspin as well as matrix-metallo-proteinases -1 and -9 as well as vascular adhesion molecule endoglin are interesting candidates for future research.