Abstract 1957: MicroRNA analysis paired with a novel live cell viability assay: a complete epigenetic workflow in human cancer cell lines

The requirements for nucleic acid purification for use in RNA profiling have expanded with the growing interest in the role of microRNAs (miRNAs) and other small non-coding RNAs in cancer cell growth and metastasis. This evolution of expression analysis has highlighted the need for more sophisticated tools for total RNA isolation from cancer cells and tissues, beyond traditional translated messenger RNAs (mRNA). To address this need, we describe tools to isolate total RNA, for mRNA and miRNA analyses, in the context of an epigenetic workflow with cancer cell lines. In these experiments, histone deacetylase (HDAC) inhibitor treatment of different cancer cell lines was performed and expression of miRNA and mRNAs measured in parallel with assays for cell viability, cytotoxicity, and HDAC activity. This workflow multiplexes RNA and miRNA expression profiling with assays for viability and cytotoxicity to efficiently capture a more complete understanding of expression results. With this improved cell viability workflow, one experiment from the same well can yield information about cell health, cytotoxicity, HDAC activity, and mRNA plus miRNA expression profiling. Our workflow incorporates monitoring of cell health in real-time during experimental treatments, simple and effective total RNA purification, and robust quantitative PCR reagents for a comprehensive experimental design. Citation Format: Samantha Lewis, Brad Hook, Don Smith, Douglas Horejsh, Trista Schagat. MicroRNA analysis paired with a novel live cell viability assay: a complete epigenetic workflow in human cancer cell lines. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1957.