Mutation detection in the interaction domains of NRF2 and KEAP1 in lung cancer cell lines / 重庆医学

ABSTRACT

Objective To detect the interaction domains' gene mutations of KEAP1 and NRF2 in non-small-cell lung cancer cell lines,and to analyze the significance of these mutations on the study of lung cancer cell lines.Methods Six non-small-cell lung cancer cell lines were used as the research materials.The 2nd to 6th exons of KEAP1 and the coding sequences of DLG and ETGE motif were amplified by PCR,and the products were used for gene sequencing analysis.Obtained gene sequences were analyzed using NCBI databases to get the base locations of gene mutations,as well as the subsequent amino acid sequence changes.Meanwhile,the relative intracellular reactive oxygen species (ROS) concentrations in the tested cell lines were detected and used in the analysis of abnormal NRF2 transcriptional activity in lung cancer cell.Results Four mutations were detected in the 4th exon of KEAP1 gene from all the 6 cancer cell lines,several other missense mutations were also investigated in the 3rd exon of KEAP1 from some cancer cell lines.Multiple genetic variations were found in all the NRF2-ETGE motif-encoding sequences of the 6 cancer cell lines.All 6 cancer cell lines were found to have higher ROS concentration than the lung germ cell line.Conclusion Lung cancer cells generally contain high levels of ROS as well as gene mutations in KEAP1 and NRF2 genes which lead to abnormal transcriptional activity of NRF2 in lung cancer cell lines.