Analysis of Kras2 gene mutations and new generation sequencing (NGS) on candidate chromosomal regions associated with susceptibility/resistance to inflammatory response

Abstract

Mice AIRmax (high response) and AIRmin (low response) mice produced by bidirectional genetic selection based on the intensity of acute inflammatory response (AIR) differ significantly for other phenotypes such as tumor development susceptibility. The AIRmax are very resistant and AIRmin are very susceptible to chemically induced of skin and lung tumors, indicating common gene regulation between the phenotypes of acute inflammatory reactivity and predisposition toward carcinogenesis. The AIRmax and AIRmin mice showed, respectively, the haplotypes of the Pas1 locus resistance and susceptibility, respectively, which is the major genetic factor involved in the susceptibility to lung in mice. This may indicate the participation of this region (which contains the oncogene Kras2), in the regulation of acute inflammatory response. Our proposal is to identify Kras2 activating mutations using pirosequencing assay, and gene expression levels of Kras4A and Kras4B isoforms by qPCR in lung tumors of mice from the F2 population (AIRmax x AIRmin), resulting from the interbreeding of two lineages. Then these date will be used as phenotypes in linkage analysis with SNPs (GWA for genome wide association) in order to identify genetic elements regulating the mutability and oncogene expression (AU)