Oxidative Stress-Induced Mitochondrial DNA Methylation Affect Cell Viability
Date Issued
2012
Date
2012
Author(s)
Yang, Chih-Hsin
Abstract
Epigenetics which is defined as ” A phenomenon that changes the phenotype without changing the underlying DNA sequence, and it can inherit to offspring.” is one of the major research topics in modern biology. In addition, epigenetics includes DNA methylation, histone modification, and non-coding RNA. In which DNA methylation is associated with gene imprinting, embryonic development, X-chromosome inactivation, oncogenesis, and gene silencing. By focusing on DNA methylation, there are lots of researches in nuclear DNA, however, the information about mitochondrial DNA methylation is relatively rare. Nevertheless, it is known that mitochondria play an important role in ATP producing.
Previous study shows a low methylation pattern in mitochondria and consider that the relationship between mtDNA methylation and cell viability is weak . But, there were two papers which were published in 2009 and 2011 respectively were focusing on this issue as well. One indicates the different methylated accessibility of mtDNA and the other shows the appearance of mitochondrial DNA methyltransferase 1. Recently, the mitochondrial DNA methylation might need re-evaluation due to the detection technology progress.
In this project, one of the major technology, pyrosequencing which is more direct and accurate method to detect oxisative stress-induced mtDNA methylation on HeLa cell and PC9 cell, is applied. By carrying out this technique, five mitochondrial-encode gene: ATP6, ND1, ND6, TERM and D-loop are analysed.
Based on our research, PC9 cell has better viability than HeLa cell after H2O2 treatment. Moreover, methylation level of five genes are all increasing in PC9 cell, but no significant difference in HeLa cell. Therefore, baseline data shows that HeLa cell has higher methylation level than PC9 in mtDNA, however, the methylation level of PC9 cell is above than HeLa cell after H2O2 treatment. It is believed that oxidative stress-induced DNA methylation has cell-specificity and the methylation level might affect cell viability.
Subjects
mitochondrial DNA
methyltransferase
methylation
oxidative stress
Type
thesis
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