The class 3 semaphorins, sema3A and sema3C, provide important guidance cues in cell development and in cancer; however, the role of these semaphorins in prostate cancer is not known. We report here that sema3A transfected cells exhibit decreased invasion and adhesion in Matrigel-based assays and that sema3C transfected cells exhibit increased invasive and adhesive characteristics. Important adhesion proteins were differentially modulated in sema3A and sema3C cells in a manner consistent with their subsequent invasive and adhesive characteristics. E-cadherin expression as determined by Western blot analysis was strongly upregulated in sema3A transfected cells, but strongly downregulated in sema3C transfected cells compared to untransfected and mock empty vector-transfected PC-3 cells. ß-catenin levels were not changed in sema3A transfected cells; however, sema3C transfected cells had lower expression of this protein. Sema3C transfected cells exhibited greater cellular membrane expression of certain • integrins as compared to untransfected and sema3A transfected cells, a characteristic associated with increased adhesion and invasion. These data indicate that the invasive ability of sema3A and sema3C transfected PC-3 cells is, in part, correlated with adhesion protein expression and adhesive ability to constituents of neighboring cells and the extracellular matrix.

Resveratrol has been shown to have anticarcinogenic activity. We previously found that resveratrol inhibited growth and induced apoptosis in 2 human melanoma cell lines. In this study we determined whether resveratrol would inhibit human melanoma xenograft growth. Athymic mice received control diets or diets containing 110 micromol/L or 263 micromol/L resveratrol, 2 wk prior to subcutaneous injection of the tumor cells. Tumor growth was measured during a 3-wk period. Metabolism of resveratrol was assayed by bolus gavage of 75 mg/kg resveratrol in tumor-bearing and nontumor-bearing mice. Pellets containing 10-100 mg resveratrol were implanted into the mice, next to newly palpated tumors, and tumor growth determined. We also determined the effect of a major resveratrol metabolite, piceatannol, on experimental lung metastasis. Resveratrol, at any concentration tested, did not have a statistically significant effect on tumor growth. The higher levels of resveratrol tested (0.006% in food or 100 mg in slow-release pellets) tended to stimulate tumor growth (P = 0.08-0.09). Resveratrol and its major metabolites, resveratrol glucuronide and piceatannol, were found in serum, liver, skin, and tumor tissue. Piceatannol did not affect the in vitro growth of a murine melanoma cell line, but significantly stimulated the number of lung metastases when these melanoma cells were directly injected into the tail vein of the mouse. These results suggest that resveratrol is not likely to be useful in the treatment of melanoma and that the effects of phytochemicals on cell cultures may not translate to the whole animal system.

Androgen-independent prostate cancer is resistant to therapy and is often metastatic. Here we studied the effect of deprivation of tyrosine and phenylalanine (Tyr/Phe), glutamine (Gln), or methionine (Met), in vitro on human DU145 and PC3 androgen-independent prostate cancer cells, and on nontumorigenic human infant foreskin fibroblasts and human prostate epithelial cells. Deprivation of the amino acids similarly inhibited growth of DU145 and PC3 cells, arresting the cell cycle at G0/G1. Met and Tyr/Phe deprivation induces apoptosis in DU145, but only Met deprivation induces apoptosis in PC3 cells. The growth of normal cells is inhibited, but no apoptosis is induced by amino acid deprivation. Tyr/Phe deprivation inhibits expression and phosphorylation of focal adhesion kinase (FAK) and extracellular-regulated kinase (ERK) in DU145 but not PC3 or normal cells. Met deprivation inhibits phosphorylation but not protein expression of FAK and ERK in PC3. Therefore, apoptosis of DU145 and PC3 cells by amino acid restriction is FAK and ERK dependent. Tyr/Phe and Met deprivation inhibits invasion of DU145 and PC3, but Gln deprivation only inhibits invasion of DU145 cells. This indicates that the inhibition of invasion is not dependent on induction of apoptosis. The inhibition of invasion by Tyr/Phe restriction in DU145 and Met restriction in PC3 is consistent with the inhibition on FAK/ERK signaling. The inhibition of Tyr/Phe restriction in PC3 and Gln restriction in DU145 is not associated with inhibition of FAK/ERK. This indicates that FAK/ERK-dependent and independent pathways are modulated by specific amino acid restriction. This study shows the potential for specific amino acid restriction to treat prostate cancer.

This study examined the mechanism underlying the increase of peripheral memory phenotype T cells that occurs during chronic alcohol consumption in mice. Female C57BL/6 mice were given 20% (w/v) alcohol in the drinking water for 2 weeks to 6 months. Chronic alcohol consumption significantly induced peripheral T cell lymphopenia; up-regulated expression of CD44 on T cells and increased the percentage of CD4+CD44int/hi and CD8+CD44int/hi Ly6C+ T cells; up-regulated the expression of CD43 on CD8+ T cells; increased the percentage of interferon--producing T cells; decreased the percentage of CD8+CD28+ T cells; and down-regulated the expression of CD28 on CD4+ T cells. Expression of CD25 and CD69 on peripheral CD8+ T cells was not affected and inconsistently expressed on CD4+ T cells. Neither cell type showed altered expression of CD137 or CD153. Alcohol withdrawal did not abrogate the increase in CD8+Ly6C+cells induced by alcohol consumption. In vivo bromodeoxyuridine incorporation experiments demonstrated that chronic alcohol consumption decreases naïve T cells that are presumed to have emigrated from the thymus and increases proliferation of memory T cells, but accelerates peripheral T cell turnover. Together these results indicate that chronic alcohol consumption results in T cell lymphopenia, which in turn induces T cell homeostatic proliferation that increases the proportion of peripheral memory T cells relative to naïve T cells.

The role of β-endorphin (β-EP) in ethanol-altered NK cell cytolytic activity is studied using male Fischer-344 rats as an animal model. Ethanol was administered for 1, 2, 3, or 4 wk in a liquid diet containing 8.7% ethanol (v/v), which means that 37% of the total calories were derived from ethanol. Rats treated with ethanol for 1 wk showed an increase in hypothalamic and plasma levels of immunoreactive (IR)-β-EP, but displayed no significant effect on NK cell activity determined by 51Cr release assay, as compared with those in pair-fed and ad libitum-fed animals. However, animals treated with ethanol for 2, 3, or 4 wk showed decreased hypothalamic and plasma levels of IR-β-EP and decreased splenic NK cell activity. No significant decrease in the number of splenocytes and NK cells or in the percentage of NK cells was seen until after 3 and 4 wk of ethanol treatment. Exposure in vitro of splenic lymphocytes obtained from control animals to various concentrations of β-EP increased NK cell activity. The opiate antagonist naltrexone blocked the β-EP-stimulated effect. The in vitro NK cell response to β-EP was reduced in the splenocytes obtained from animals treated with ethanol for 2 wk, but not in those obtained from animals treated with ethanol for 1 wk as compared with those in control animals. Additionally, β-EP administration into the paraventricular nucleus of the hypothalamus stimulated NK cell cytolytic activity, whereas the opiate blocker administration reduced NK cell activity. The NK cell responses to paraventricular nucleus β-EP were reduced in the animals treated with ethanol for 2 wk. These data provide evidence for the first time that ethanol inhibits NK cell cytolytic activity, possibly by reducing β-EP-regulated splenic NK cell function.