Accumulation of somatic mutation in mitochondrial DNA extracted from peripheral blood cells in diabetic patients.
OBJECTIVE
A point mutation of mitochondrial DNA at nucleotide number 3243 A to G is responsible for both the major genetic aetiologies of the MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes) and mitochondrial diabetes. Otherwise, this mutation is also reported to occur as an acquired somatic mutation, possibly due to oxidative stress. Since diabetes can cause severe oxidative stress, we hypothesize that the accumulation of the somatic 3243 A to G mutation in mitochondrial DNA can be accelerated by diabetes.


METHODS
DNA was extracted from blood samples of 290 non-diabetic healthy subjects (age 20-60) including 98 newborn infants and from 383 patients with Type II (non-insulin-dependent) diabetes mellitus (age 18-80). The extent of somatic 3243 A to G mutation to total mitochondrial DNA was detected by real-time PCR using the TaqMan Probe.


RESULTS
Whereas the level of the 3243 A to G mutation was negligible in the newborn group, it was increased in healthy subjects who were 20 to 29 and 41 to 60 years of age, suggesting that this mutation was somatic. In the diabetic patients the mutation rate increased along with age and the duration of diabetes. In the middle-aged group (age 41-60), the 3243 A to G mutation accumulates fourfold higher in the diabetic patients than the healthy subjects. Moreover, multiple regression analysis showed that the most critical factor associated with this mutation in diabetic patients was the duration of diabetes.


CONCLUSIONS
Diabetes accelerates the accumulation of the somatic 3243 A to G mutation in mitochondrial DNA, which can accelerate the ageing process. This somatic mutation could possibly be a new marker for estimating the duration of diabetes.
Nomiyama
T
T
Department of Medicine, Metabolism and Endocrinology, School of Medicine, Juntendo University, Tokyo, Japan.
Tanaka
Y
Y
Hattori
N
N
Nishimaki
K
K
Nagasaka
K
K
Kawamori
R
R
Ohta
S
S
D000225:Adenine; D000293:Adolescent; D000328:Adult; D017668:Age of Onset; D000368:Aged; D001483:Base Sequence; D015992:Body Mass Index; D004247:DNA / Q000097:blood* / Q000235:genetics; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics*; D003924:Diabetes Mellitus, Type 2 / Q000097:blood / Q000235:genetics*; D005260:Female; D006147:Guanine; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D017354:Point Mutation; D012016:Reference Values
D017931:DNA Primers; D004272:DNA, Mitochondrial; D006147:Guanine; D004247:DNA; D000225:Adenine