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Introduction: Incurable metastasis still remains the major cause of death among women diagnosed with breast cancer. There are meanwhile comprehensive molecular profiles available from metastatic breast cancer, providing huge lists for novel candidate genes for metastatic spread. The function of most of these genes is unknown. Recently, miRNAs have been added as potential crucial regulators of metastatic spread, and corresponding profiles from metastatic breast cancer are available as well. Within this project, a set of the 100 most promising genes and miRNAs were selected from these molecular profiles to study, which of these promote or prevent spreading of the cancer cells. Purpose: We selected a set of 100 candidate genes and miRNAs from available molecular profiles of metastatic breast cancer to produce isogenic stable cell lines in order to identify new crucial regulators of metastasis formation. Material: MCF7 breast cancer cell line, vectors for stable integration and recombination, library of gateway cloned miRNAs and genes, cell titer blue (viability assay), matrigel invasion assay. Methods: In previous work we have established a unique recombinationbased system, which allows the serial introduction of genes into cancer cell lines, including MCF7 breast cancer cells. This technique creates standardized stable cell line libraries with over expression of a gene in a tetracycline-inducible fashion. These cell lines are subjected to serial cell viability and invasion assays. Results: Having analyzed a subset of the candidate genes by cell viability assays, we so far identified a number of genes that modulate basic breast cancer growth. Conclusions: Employing our recombination-based system allows for rapid serial analysis of cancer candidate genes and resulted so far in the identification of novel breast cancer growth modulators. Studies of invasion are in progress.
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