Comparative analysis of Adam33 mutations in murine lung cancer cell lines by droplet digital PCR, real-time PCR and Insight Onco™ NGS

Abstract

In a mouse-based carcinogen bioassay, we can identify lung cancer-specific oncogenic driver mutations in circulating cell-free DNA in the blood prior to autopsy. These mutations could be used as an early biomarker for lung cancer. We investigated single nucleotide variants in gDNA isolated from LA-4 and KLN205 cell lines, through whole exome sequencing. SNVs of 15 representative genes related to lung cancer (such as Adam33) were confirmed. Among them, Adam33 increased the risk of chronic obstructive pulmonary disease (COPD), which is reported to be associated with lung cancer. Therefore, we selected Adam33, an asthma-related gene, for this study. The sensitivity of real-time PCR, droplet digital PCR and Insight Onco™ NGS to detect rare mutations of Adam33 was 10%, 1%, and 0.05%, respectively. We confirmed that the Insight Onco™ technique is a sensitive method that can be used to detect lung-specific rare mutations in vitro.