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《Chinese Journal of AIDS & STD》 2010-02
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Detection of NRTIs-induced mitochondrial DNA depletion using real-time fluorescence quantitative PCR

SUN Yu, ZHANG Hong-hai, SHI Ying, et al. (Research Laboratory of STD / AIDS, Beijing You An Hospital, Capital University of Medical Science, Beijing 100069, China)  
Objective Using real-time fluorescence quantitative PCR to detect mitochondrial DNA content changes within HepG2 cells induced by d4T and AZT. Methods HepG2 cells were treated with different concentrations(0,3,10,100,200,300μmol/L) of d4T and AZT respectively for two weeks. And then mitochondrial DNA contents were detected by real-time fluorescence quantitative PCR. Results Real-time fluorescence quantitative PCR was set up successfully to detect mitochondrial DNA contents. Mitochondrial DNA relative amounts were 96.94±5.77, 53.73±7.14, 20.78±3.10, 1.37±0.29 respectively with d4T concentrations of 0, 3, 10, 100μmol/L. The differences between groups were significant(P0.01). However, they were 96.94±5.77, 108.84±7.80, 172.56±4.70, 199.51±10.37, 158.74±6.64 and 64.06±6.27 respectively with AZT concentrations of 0, 3, 10, 100, 200, 300μmol/L, and the differences between groups were significant(P0.01). Conclusions It is practicable to detect mitochondrial DNA contents with real-time fluorescence quantitative PCR. It is d4T ,rather than AZT , that causes a dose-dependent mitochondrial DNA depletion within HepG2 cells.
【Fund】: 国家科技部“十一五”重大专项(2008ZX10001-007);; 国家自然科学基金(30870853)~~
【CateGory Index】: R450
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