Abstract 1515: Simultaneous detection of KRAS and TP53 mutations in human cancer cell lines using multiplex qPCR

Genetic alterations in KRAS and TP53 genes are common in many cancers and the understanding of their role in progression of cancer is vital for the design of successful treatment approaches. Studies suggest that mutations in KRAS and TP53 genes independently hindered the use of EGFR inhibiting drugs like Cetuximab (Erbitux) for the treatment of colorectal cancer (1). The genetic alterations in these genes are now routinely checked before starting any expensive treatments. Here, we successfully demonstrated use of duplex mutli-color SNP genotyping assays to detect KRAS and TP53 mutation in human colon cancer cell lines. This method can be potentially used to analyze samples with limited quantity and get as much information as possible from a single sample. To study the impact of these mutations, we generated gene expression profiles of 672 genes related to signaling pathways by using the Human signal transduction medium-throughput OpenArray® Panel. Selected cancer genes like BRAC1, BAX, CFLAR, MYC and others were evaluated using multiplexed single-tube PCR simultaneously detecting four different targets in the same sample. These results demonstrate that duplex SNP genotyping and multiplexed gene expression profiling from a single sample increases research capacity in situations when sample is limited. Reference: 1. A Oden-Gangloff, F Di Fiore, F Bibeau, A Lamy, G Bougeard, F Charbonnier, F Blanchard, D Tougeron, M Ychou, F Boissière, F Le Pessot, J-C Sabourin, J-J Tuech, P Michel and T Frebourg.TP53 mutations predict disease control in metastatic colorectal cancer treated with cetuximab-based chemotherapy. British Journal of Cancer (2009) 100, 1330-1335 Citation Format: Sunali Patel, Cora Woo, Joyce Wilde, Sundiep Phanse. Simultaneous detection of KRAS and TP53 mutations in human cancer cell lines using multiplex qPCR. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1515. doi:10.1158/1538-7445.AM2014-1515