Apparent Polyploidization after Gamma Irradiation: Pitfalls in the Use of Quantitative Polymerase Chain Reaction (qPCR) for the Estimation of Mitochondrial and Nuclear DNA Gene Copy Numbers

被引：3

作者：

Kam, Winnie W. Y. ^{[1
,2
]}

Lake, Vanessa ^{[1
]}

Banos, Connie ^{[1
]}

Davies, Justin ^{[1
,3
]}

Banati, Richard ^{[1
,2
,4
]}

机构：

[1] Australian Nucl Sci & Technol Org, Sydney, NSW 2234, Australia

[2] Univ Sydney, Fac Hlth Sci, Sydney, NSW 2141, Australia

[3] Univ Sydney, Sch Phys, Sydney, NSW 2006, Australia

[4] Univ Sydney, Natl Imaging Facil, Brain & Mind Res Inst, Sydney, NSW 2050, Australia

来源：

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2013年 / 14卷 / 06期

关键词：

gamma radiation; temperature; mitochondria; nucleus; DNA copy number; polyploidization; ACUTE LYMPHOBLASTIC-LEUKEMIA; DOUBLE-STRAND BREAKS; IONIZING-RADIATION; SUPEROXIDE-DISMUTASE; OXIDATIVE DAMAGE; CELL-LINE; COMPLEX-I; TEMPERATURE; PCR; QUANTIFICATION;

D O I：

10.3390/ijms140611544

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Quantitative polymerase chain reaction (qPCR) has been widely used to quantify changes in gene copy numbers after radiation exposure. Here, we show that gamma irradiation ranging from 10 to 100 Gy of cells and cell-free DNA samples significantly affects the measured qPCR yield, due to radiation-induced fragmentation of the DNA template and, therefore, introduces errors into the estimation of gene copy numbers. The radiation-induced DNA fragmentation and, thus, measured qPCR yield varies with temperature not only in living cells, but also in isolated DNA irradiated under cell-free conditions. In summary, the variability in measured qPCR yield from irradiated samples introduces a significant error into the estimation of both mitochondrial and nuclear gene copy numbers and may give spurious evidence for polyploidization.

引用

页码：11544 / 11559

页数：16