Circadian variation in O6-alkylguanine-DNA alkyltransferase activity in circulating blood mononuclear cells of healthy human subjects.
Cytotoxic agents such as chloroethylnitrosoureas (CENUs) mostly alkylate DNA on the O6-guanine position. This highly mutagenic lesion can be repaired by O6-alkylguanine-DNA alkyltransferase (AGT), which removes the alkyl group by accepting it to the cysteine residue of its active site. AGT activity displayed a circadian rhythm in mouse liver, coincident with that of CENU tolerability. We investigated whether AGT activity displayed a circadian rhythm in human circulating mononuclear cells (MNCs). The study was performed in 12 healthy volunteers aged 19 to 31 years. Circadian synchronization was verified with rest/activity cycle as assessed with wrist actigraphy and plasma cortisol and melatonin rhythms. Subjects were hospitalized for 24 hr and blood samples were obtained at 08:00, 12:00, 16:00, 20:00, 22:00, 00:00, 02:00, 04:00 and 08:00 overnight. MNCs were isolated on Ficoll immediately after blood sampling and frozen at -196 degrees C until AGT activity determination by HPLC. Mean AGT activity (+/- SEM) varied from 821 +/- 67 fmol/mg of total proteins at noon (trough), up to 1,055 +/- 80 fmol/mg at midnight (peak), i.e., by approximately 30%. A circadian rhythm was statistically validated with both analysis of variance (p < 0.009) and cosinor (p < 0.02) for AGT activity in MNCs (acrophase +/- SD at 00:30 +/- 210 min) as well as for MNC circulating count and for plasma cortisol and melatonin concentrations. Despite individual variations in the extent of AGT activity rhythm (more or less pronounced according to subject), AGT activity displayed a circadian rhythm in human MNCs of our healthy study group subjects. The results warrant to further investigate AGT rhythmicity both in circulating MNCs and in target tissues of cancer patients, as a prerequisite for clinical testing of chronotherapy with alkylating agents.
Marchenay
C
C
INSERM U484, Clermont-Ferrand, France. Recherche.Clinique@cjp.u-clermont1.fr
Cellarier
E
E
Lévi
F
F
Rolhion
C
C
Kwiatkowski
F
F
Claustrat
B
B
Madelmont
J C
JC
Chollet
P
P
D000328:Adult; D000704:Analysis of Variance; D002851:Chromatography, High Pressure Liquid; D002940:Circadian Rhythm; D006801:Humans; D006854:Hydrocortisone / Q000097:blood; D007963:Leukocytes, Mononuclear / Q000201:enzymology*; D008297:Male; D008550:Melatonin / Q000097:blood; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000097:blood*; D013997:Time Factors
D019853:O(6)-Methylguanine-DNA Methyltransferase; D008550:Melatonin; D006854:Hydrocortisone