Worldwide oral squamous cell carcinoma (OSCC) represents about 5.5% of all malignancies, with f30,000 new cases each year in the United States. The integrin A v B 6 and the enzyme cyclooxygenase-2 (COX-2) are implicated in OSCC progression and have been suggested as possible therapeutic targets. Each protein also is reported to identify dysplasias at high risk of malignant transformation, and current clinical trials are testing the efficacy of nonsteroidal anti-inflammatory drugs (NSAID) at preventing OSCC development. Given the probable increased expression of A v B 6 and COX-2 in OSCC and the inhibition of several integrins by NSAIDs, we investigated whether NSAIDs affected A v B 6 -dependent cell functions. We found that expression of both A v B 6 and COX-2 was significantly higher in OSCC compared with oral epithelial dysplasias. Neither protein preferentially identified those dysplastic lesions that became malignant. Using OSCC cell lines, modified to express varying levels of A v B 6 , we assessed the effect of COX-2 inhibition on cell invasion. We found that the COX-2 inhibitor NS398 inhibited specifically A v B 6 -dependent, but not A v B 6 -independent, OSCC invasion in vitro and in vivo, and this effect was modulated through prostaglandin E 2 (PGE 2 )-dependent activation of Rac-1. Transient expression of constitutively active Rac-1, or addition of the COX-2 metabolite PGE 2 , prevented the anti-invasive effect of NS398. Conversely, RNA interference down-regulation of Rac-1 inhibited A v B 6 -dependent invasion. These findings suggest that COX-2 and A v B 6 interact in promoting OSCC invasion. This is a novel mechanism that, given the ubiquity of A v B 6 expression by head and neck cancers, raises the possibility that NSAIDs could protect against OSCC invasion. (Cancer Res 2006; 66(22): 10833-42)

SummaryMany animal species exhibit sex differences in aging. In the nematode Caenorhabditis elegans , under conditions that minimize mortality, males are the longer-lived sex. In a survey of 12 independent C. elegans isolates, we find that this is a species-typical character. To test the hypothesis that the C. elegans male longevity bias evolved as a consequence of androdioecy (having males and hermaphrodites), we compared sex-specific survival in four androdioecious and four dioecious (males and females) nematode species. Contrary to expectation, in all but C. briggsae (androdioecious), males were the longerlived sex, and this difference was greatest among dioecious species. Moreover, male lifespan was reduced in androdioecious species relative to dioecious species. The evolutionary theory of aging predicts the evolution of a shorter lifespan in the sex with the greater rate of extrinsic mortality. We demonstrate that in each of eight species early adult mortality is elevated in females/ hermaphrodites in the absence of food as the consequence of internal hatching of larvae (matricide). This age-independent mortality risk can favour the evolution of rapid aging in females and hermaphrodites relative to males.

A number of recent studies of aging in Drosophila, mice and dogs have shown an association between reduced body size and increased lifespan. It is unclear (a) whether such an association is a general feature of animal species; and (b) whether the association reflects an effect of body size on aging, or pleiotropic effects of common determinants of growth and aging. To address these issues, we have studied the relationship between size and lifespan in the nematode Caenorhabditis elegans, and surveyed related findings in Drosophila. In C. elegans, we compared 12 wild isolates with varying body size and lifespan, but saw no correspondence between these traits. We also examined aging in giant and dwarf mutants, but observed only reduced lifespan in all cases. In a comparison of 15 long-lived daf-2 insulin/IGF receptor mutants, we saw a positive correlation between body length and lifespan, and up to a 28% increase in daf-2 mutant body volume. Thus, in C. elegans, insulin/IGF signaling may limit growth rather than promote it. Studies of Drosophila show no consistent correlation between body size and lifespan. These results indicate that the negative correlation between body size and lifespan seen in some mammals is not typical of invertebrates, but support the view that co-variation of size and longevity may occur via effects on insulin/IGF signaling. q

Learning is an adaptive change in behavior in response to environmental stimuli. In mammals, there is a distinct female bias to learn skills that is still unprecedented in other animal taxa. Here we have investigated the biological determinants of performance in an associative learning paradigm in the nematode Caenorhabditis elegans. Using an assay of chemotactic reactions associated with food deprivation, wild-type male worms show inferior learning ability relative to hermaphrodites. Sex-based learning difference is therefore an ancient evolutionary feature appearing even in relatively simple animals. C. elegans mutants with reduced insulin/IGF-1 signaling also exhibit a greatly reduced learning ability in this assay. In addition, hyperactivation of insulin/IGF-1 signaling through loss-of-function mutations in the PTEN phosphatase daf-18, a negative regulator of insulin/IGF-1 signaling, enhances learning ability beyond that of wild type. According to our epistasis analysis, the effect of DAF-2 on learning acts via phosphatidylinositol 3,4,5-trisphosphate (PIP 3 ) production, but not the DAF-16 FOXO transcription factor. This implies that the signaling pathway from DAF-2 affecting this learning paradigm branches between PIP 3 production and DAF-16. However, learning capacity of nematodes is lowered by loss-offunction mutations in daf-16, suggesting involvement of noninsulin/IGF-1 signaling-dependent DAF-16 activation in learning. Potentially, sex and insulin/IGF-1 signaling affect performance in this learning assay via effects on the neurobiology of learning.

Sex differences in longevity and aging are seen throughout the animal kingdom. These are likely to result, in part, from sex differences in endocrinology. In the nematode Caenorhabditis elegans, males are the longer-lived sex. Here we explore the possibility that sex differences in insulin/insulin-like growth factor 1 (IGF-1) and steroid endocrinology contribute to this sex difference in aging by studying C. elegans populations in liquid culture. We report that in hermaphrodite populations, mutational loss of the DAF-12 steroid receptor affected life span as in previous plate-culture studies: mutant longevity is suppressed in a weak daf-2 insulin/IGF-1 receptor mutant but enhanced in a stronger daf-2 mutant. However, in males, mutation of daf-12 had little effect on aging in either weak or strong daf-2 mutants. Moreover, while mutation of daf-12 marginally reduced life span in daf-2(+) hermaphrodites, as in plate-cultured populations, it did not in daf-2(+) males. These results could imply that in C. elegans, as in mammals, sex differences in steroid endocrinology contribute to sex differences in aging.