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28 June 2012

Altered expression of base excision repair genes in response to high glucose-induced oxidative stress in HepG2 hepatocytes

Jing PangABCDEFG, Chao XiBCEFG, Yang DaiBC, Huan GongBC, Tie-mei ZhangADEFG

DOI: 10.12659/MSM.883206

Med Sci Monit 2012; 18(7): BR281-285

Abstract

Background: It is widely accepted that chronic hyperglycemia induces DNA oxidative damage in type 2 diabetes, but little is known about the effect of hyperglycemia on the DNA repair system which plays a critical role in the maintenance of genomic DNA stability in diabetes.
Material/Methods: To investigate the alteration of base excision repair (BER) genes under hyperglycemia, the relative expression of the mRNAs of the BER genes – ogg1, polbeta, lig3, xrcc1, and parp1 – were quantified using real-time PCR in HepG2 hepatocytes incubated with 5.5 mM or 30 mM glucose.
Results: High levels of glucose induced ROS accumulation and DNA damage, paralleling the dynamic alterations of BER mRNA expression. Compared to 5.5 mM glucose-treated cells, ogg1 and polbeta mRNA expression transiently increased at day 1 and decreased after day 4 in cells exposed to 30 mM glucose. Exposure to 30 mM glucose increased the activity of PARP1, which led to reduced cellular NAD content and insulin receptor phosphorylation.
Conclusions: Exposure to high concentrations of glucose initially led to the increased expression of BER mRNAs to counteract hyperglycemia-induced DNA damage; however, long-term exposure to high glucose concentrations reduced the expression of mRNA from BER genes, leading to accumulated DNA damage.

Keywords: RNA, Messenger - metabolism, Poly(ADP-ribose) Polymerases - metabolism, Phosphorylation - drug effects, Oxidative Stress - genetics, NAD - metabolism, Insulin - pharmacology, Hepatocytes - metabolism, Glucose - pharmacology, Gene Expression Regulation, Neoplastic - drug effects, Enzyme Activation - drug effects, DNA Repair - genetics, DNA Damage - genetics, Reactive Oxygen Species - metabolism, Receptor, Insulin - metabolism, Transcription, Genetic - drug effects

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750