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Mitochondrial encoding ATPase genome damage in intestinal epithelial cells induced by hydrogen peroxide

Objective To investigate the injury effects of hydrogen peroxide on construction and function of mitochondrial DNA (mtDNA) encoding ATPase gene in intestinal epithelial cells. Methods Intestinal epithelial cells SW-480 were treated with saline solution (control) or hydrogen peroxide (H 2O 2), a representative ROS. mtDNA was extracted from the intestinal epithelial cells treated with 4 mmol/L H 2O 2. Polymerase chain reaction (PCR) with different primers of ATPase subunits 6 and 8 (ATPase 6 and ATPase 8) was performed. The products of PCR were directly sequenced. The activities of the enzyme were determined at the same time. Results There were large-scale point mutations from 8385 to 8543 of ATPase 8. H 2O 2 could result in mutagenesis (point and deletion mutations) in mtDNA encoding ATPase 8 and significantly decrease the activity of ATPase. Conclusions H 2O 2 can significantly damage the genome of ATPase encoded by mtDNA, which may result in activity decrease of ATPase