Eric Lok

Soy Isoflavones Modulate Azoxymethane-Induced Rat Colon Carcinogenesis Exposed Pre- and Postnatally and Inhibit Growth of DLD-1 Human Colon Adenocarcinoma Cells by Increasing the Expression of Estrogen Receptor-β

We studied the effects of lifetime exposure to dietary soy isoflavones in an azoxymethane (AOM)-induced rat colon cancer model. Male pups born to Sprague-Dawley rats exposed (including during pregnancy and lactation) to soy isoflavones at either no (0 mg = control), low (40 mg), or high (1000 mg) doses/kg diet were weaned and continued receiving their respective parental diets until the end of the study. Weaned rats received 2 subcutaneous injections (15 mg/kg body weight) of AOM 1 wk apart. After 26 wk, rats were killed and the coordinates of colon aberrant crypt foci (ACF) and tumors were determined. Expression of estrogen receptor (ER)-beta was assessed in rat colon tumors and in DLD-1 human colon adenocarcinoma cells exposed to soy isoflavones. Compared with the control, soy isoflavones did not affect incidences or multiplicities of colon ACF or tumors. Low-dose soy isoflavones decreased tumor burden and size compared with the control (P < 0.05). Expression of ERbeta increased in colon tumors of soy isoflavone-treated groups compared with the control. Soy isoflavones dose-dependently arrested the growth of DLD-1 cells and at subcytotoxic levels increased the expression of ERbeta. Our results suggest that pre- and postnatal exposure to dietary soy isoflavones suppresses the growth of colon tumors in male rats. The overexpression of ERbeta in both rat colon tumors and DLD-1 cells caused by soy isoflavones suggests that ERbeta is a critical mediator in mitigating its cancer-preventive effects.

Dietary fats modulate methylmercury-mediated systemic oxidative stress and oxidative DNA damage in rats.

We examined the effects of dietary fats on methylmercury (MeHg)-induced systemic oxidative stress and oxidative DNA damage in liver and kidney of male Sprague-Dawley rats. Rats were treated with a casein-based purified isocaloric diet containing 15% by weight soy oil, docosahexaenoic acid (DHA), seal oil, fish oil, or lard for 28 days, and then gavaged with 0, 1, or 3 mg MeHg/kg BW/day for 14 days. Urine was analyzed for 8-hydroxydeoxyguanosine (8-OHdG) and isoprostane, and serum for total antioxidant capacity (TAC). Liver and kidney were analyzed immunohistochemically for 8-OHdG. Both diet and MeHg showed significant main effects on some of these markers. As compared with the vehicle control, 3 mg MeHg/kg BW significantly increased urinary 8-OHdG in the lard group, urinary isoprostane in the DHA, seal oil, and fish oil groups, while significantly decreasing serum TAC in the lard and fish oil groups. In all dietary groups, 8-OHdG positive staining was located mainly in the nuclei of various cell types in liver and kidney. MeHg expressed a significant main increasing effect on 8-OHdG-positive cells in kidney. These results suggest that both dietary fats and MeHg are important mediators of systemic oxidative stress and oxidative DNA damage.

Glutathione S-transferase-placental form expression and proliferation of hepatocytes in fumonisin B1-treated male and female Sprague–Dawley rats

Fumonisin B1 (FB1), a mycotoxin produced by a common corn contaminant Fusarium moniliforme and a hepatocarcinogen in rats, has been previously suggested to act as a poor initiator, but a better promoter of gamma-glutamyltranspeptidase (GGT)-positive rat liver preneoplastic lesions. Using glutathione S-transferase-placental form (GSTP) as a more sensitive marker of initiation, we have further evaluated the initiating capacity of various doses of purified FB1 administered (a) intraperitoneally (i.p.) to male Sprague-Dawley (SD) rats for 4 days and (b) orally (PO) to male and female SD rats for 11 days. Compared to their respective controls, significant increases in GSTP-positive hepatocytes were observed in male rats administered FB1 i.p. at 10 mg/kg body weight/day for 4 days, as well as in male and female rats treated with 35 and 75 mg/kg body weight/day FB1 p.o. for 11 days. The percentage section area of liver occupied by GSTP-positive mini-foci comprising of three to 12 cells was increased significantly in male rats given 10 mg/kg FB1 i.p., or in p.o.-treated males and females with 75 mg/kg FB1. Both i.p. and p.o. FB1 treatments resulted in dose-related enhanced hepatocyte proliferation as measured by proliferating cell nuclear antigen (PCNA) labeling with significant increases in the number of PCNA-positive nuclei at the same i.p. and p.o. dose levels where the number of GSTP-positive cells were elevated. In all studies, enhanced PCNA and GSTP expression occurred at FB1 doses which, based on serum biochemical and histopathological data previously reported from our laboratory, were shown to be hepatotoxic. Therefore, our data suggest that in a manner similar to known genotoxic carcinogens, FB1 has the capacity to initiate GSTP-positive hepatocytes with their subsequent development into GSTP mini-foci at exposure levels that induce enhanced hepatocyte proliferation in response to liver toxicity. In SD rats, this occurs as early as within 4 days of i.p. treatment or 11 days of p.o. treatment.

The effect of butylated hydroxytoluene on selected immune surveillance parameters in rats bearing enzyme-altered hepatic preneoplastic lesions.

Selected immune function parameters were examined in male Fischer 344 rats following (a) induction of enzyme-altered preneoplastic liver foci (EAF), and (b) growth modulation of EAF by 30-day feeding with the food antioxidant butylated hydroxytoluene (BHT). Glutathione S-transferase-P (GSTP)-positive EAF were observed in livers of rats receiving diethylnitrosamine (DEN), 2-acetylaminofluorene (2-AAF) and partial hepatectomy (PH) (Solt-Farber procedure), with or without BHT treatment. The induction of EAF and/or 0.5% BHT treatment resulted in a significant reduction in the natural killer (NK) cell activity of splenocytes. PH did not affect NK activity significantly compared with control (no PH) rats. The concanavalin A-induced lymphoproliferative activity of splenocytes was increased in rats with PH compared with those without. A lag in time needed to attain maximum calcium release was observed only in the rats with PH compared with those without PH. None of the treatments affected the phagocytic activity of resident peritoneal macrophages. Only EAF-bearing rats without BHT treatment had increased granulocyte and monocyte levels, while the leucocyte and lymphocyte levels were reduced by the initiator DEN. but not by BHT treatment. Further investigations are necessary to determine whether the observed suppression of NK cell activity during EAF induction and growth modulation by BHT is a contributing factor in enhancement of rodent liver neoplasia by this non-genotoxic food antioxidant.

The effect of different levels of dietary α-linolenic and other fatty acids on mammary gland ductular cell proliferation in female Swiss Webster mice

Abstract In previous work we have shown that changing the fatty acid composition of a constant amount of fat in a modified AIN-76A diet affected the level of ductular cell proliferation in the mammary glands of young virgin female Swiss Webster mice. In particular, linoleic acid concentrations of 5–10% of the total fat in the diet led to variable but appreciably higher levels of proliferation than did higher levels of linoleic acid. In this report it is shown that feeding low levels of the total fat as α-linolenic acid (0–5%) resulted in a similar effect. In addition the effects of other fats including menhaden oil, were further investigated.

Effect of addition of dried healthy or diseased parsnip root tissue to a modified AIN-76A diet on cell proliferation and histopathology in the liver, oesophagus and forestomach of male Swiss Webster mice.

Umbelliferous crop plants, including the parsnip (Pastinaca sativa L.), elaborate enhanced levels of furocoumarins, including psoralens, when subjected to biotic or abiotic stress. These furocoumarins are recognized to lead to phototoxicity. In this study, the effect of these agents, which are present in diseased parsnip root tissue, on the liver and two tissues on the route of entry to the body (the oesophagus and forestomach) were investigated. Young male Swiss Webster mice were fed for approximately 30 days with modified AIN-76A diets containing 32.5% dried healthy, 32.5% apparently healthy or 32.5% fungicide-treated parsnip root tissue, and 8, 16 or 32.5% dried diseased (Phoma complanata-infected) parsnip root tissue. As controls, three modified AIN-76A diets differing in their edible starch-to-sucrose ratios (C1-C3) were administered for an equal time. Dried healthy parsnip root tissue, compared with controls, did not significantly affect any of the indices of cellular proliferation or histopathological parameters that were assessed. Histopathological examination of the oesophagus and forestomach demonstrated no significant changes as a result of feeding any of the diets containing parsnip tissue. In the liver, the highest level (but neither of the two lower levels) of dried diseased parsnip root tissue led to swelling of the cytoplasm in cells surrounding the central vein of hepatic lobules, with consequent compression of the peripheral cells. Using [3H]thymidine radioautography, a dose-related increase in cell labelling with the level of diseased parsnip root tissue was demonstrated in the liver. Compared with control diet C2 only, the extent of [3H]thymidine labelling in the liver was increased in mice receiving apparently healthy parsnip tissue; a slight, not statistically significant, increase was also noted with fungicide-treated parsnip tissue. Increased [3H]thymidine labelling with the feeding of diseased parsnip tissue was also found in the greater curvature of the forestomach and the region of the oesophageal-forestomach junction, but not at the glandular junction of the forestomach nor in the mid-oesophagus.