We investigated the association between hypercholesterolemia or serum total cholesterol level and the C825T polymorphism in the G-protein ss3 subunit gene in Japanese workers using multivariate analysis to control possible confounding factors. The study subjects were 1,452 males and 1,169 females selected from 3,834 males and 2,591 females. Hypercholesterolemia was classified as either a total serum cholesterol $5.7 mmol/L (220 mg/dL) or taking lipid-lowering medication. The statistical power of the study was estimated as 99.9% based on total cholesterol levels of previous significant studies in Caucasians and Japanese. The genotype distributions in males with hypercholesterolemia (CC = 78, CT = 154, TT = 78) and females (CC = 60, CT = 137, TT = 52) were not significantly different from normal males (CC = 280, CT = 595, TT = 267) or females (CC = 234, CT = 501, TT = 185). Allele distributions also were not significantly different in either sex. Logistic regression analysis of hypercholesterolemia data and analysis of variance, t-test and multiple regression of serum total cholesterol data demonstrated no significant relationship with the C825T genotype. Despite sufficient statistical power, this study did not demonstrate a significant influence of C825T on hypercholesterolemia or serum total cholesterol level. The targeting of this polymorphism therefore is unlikely to be beneficial in preventing hypercholesterolemia in the general Japanese population.