Reduced cell proliferation may mediate anticarcinogenic effects of caloric restriction (CR). Using heavy water ( 2 H2O) labeling, we investigated the cell proliferation response to CR in detail, including time course, effect of refeeding, and role of intermittent feeding with 5% CR. In the time-course study, 8-wk-old female C57BL/6J mice were placed on a 33% CR regimen (fed 3 times/wk) for varying durations. Compared with responses in controls fed ad libitum (AL), proliferation rates of keratinocytes, mammary epithelial cells, and T cells were markedly reduced within 2 wk of CR. In mice fed 95% ad libitum (C95, fed 3 times/wk), cell proliferation was also reduced in all tissues so that differences from 33% CR were only significant at 1 mo. In the refeeding study, mice were refed a C95 diet for varying durations after 1 mo of 33% CR. Cell proliferation rebounded to a suprabasal rate in all tissues after 2 wk of refeeding and then normalized after 2 mo, although the C95 group again exhibited lower cell proliferation than the AL group. The role of intermittent feeding was studied by comparing 33% CR and C95 animals (both fed intermittently) with animals fed isocalorically either daily or continuously by pellet dispenser. Intermittent feeding had no additive effect on 33% CR but reduced cell proliferation in all tissues at the 95% caloric intake level. In summary, the CR effect on cell proliferation is potent, rapid, and reversible in several tissues, and an intermittent feeding pattern reproduces much of the effect in the absence of substantial CR. stable isotopes; refeeding; keratinocyte; mammary; T cell CALORIC RESTRICTION (CR), defined as undernutrition without malnutrition (45), was first discovered in 1935 to extend maximal life span in rats (29). Since then, similar findings have been reported in mice, fish, flies, worms, and yeast (45). A range of 30 -70% extension of maximal life span has been achieved using variations on CR regimens (45), including both early-and adult-onset CR (44,46). CR also exerts a number of other beneficial health effects, including reduced carcinogenesis, enhanced insulin sensitivity, and reduced cardiovascular disease risk (15). The inhibitory effect of CR on carcinogenesis is of particular interest, because CR effectively inhibits spontaneous tumor formation as well as neoplasias in knockout/ transgenic models of cancer-and chemically induced tumorigenesis (18,19,45). The mechanisms by which CR extends life span and inhibits carcinogenesis remain unknown (15,19).CR could affect several steps in the multistage carcinogenesis model (18). CR may function as an anti-initiator by decreasing carcinogen activation, enhancing carcinogen detoxification, scavenging reactive oxygen species, or enhancing DNA repair (18). CR could also function as an antipromoter by reducing mitoses of initiated cells, altering expression of cancer-related genes, decreasing inflammation, enhancing immune competence, or stimulating apoptosis (18). Reductions in cell proliferation might be expected in view of ob...