Analysis of vault RNA expression level using qPCR

The project's main hypothesis is that vault RNAs are involved in cancer survival and response to therapy. Consequently, the project will answer the following questions: i) What is the basal expression level of human vault RNA paralogs in various breast cancer cell lines? ii) What is the role of vtRNAs paralogs (siRNA knockdown will be involved) in breast cancer cell proliferation and survival exposed to therapeutic agents (doxorubicin or cisplatin)? The panel of selected breast cancer cell lines involves MCF-7, MDA-MB-231, MDA-MB-453, MDA-MB-468, SK-BR-3, and T-47D, as well as non-cancer breast cells (MCF-12A). Cell line selection was based on the differences between cell genotypes and phenotypes (i.e., different invasiveness, drug resistance, growth factor receptors, etc.). After screening with RT-qPCR, three cell lines with the most abundant basal vault RNAs were selected for further experiments. Selected cell lines, MDA-MB-231, T47D and MCF12A were treated with cisplatin and doxorubicin in subcytotoxic concentrations. Finally, MDA-MB-231 cell line was a subject in the last part of the experiment regarding to the best model for this project evaluation and treated with 3 concentrations (subcytotoxic and higher) in time course 24h, 48h, 72h.