Cordyceps cicadae induces G2/M cell cycle arrest in MHCC97H human hepatocellular carcinoma cells: a proteomic study.

BACKGROUND: Cordyceps cicadae is a medicinal fungus that is often used for treating cancer. However, the anticancer mechanisms of C. cicadae are largely unknown. This study aims to investigate the anticancer mechanisms of C. cicadae against hepatocellular carcinoma cells in vitro using a proteomic approach. METHODS: Human hepatocellular carcinoma MHCC97H cells were treated with a water extract of C. cicadae (0, 100, 250, 500, and 1000 µg/mL) for 48 h and harvested for cell viability assays. The significant differences in protein expression between control and C. cicadae-treated cells were analyzed by two-dimensional gel-based proteomics coupled with matrix-assisted laser desorption ionization-time of flight mass spectrometry. Flow cytometry analysis was employed to investigate the cell cycle and cell death. The anticancer molecular mechanism was analyzed by whole proteome mapping. RESULTS: The water extract of C. cicadae (0, 100, 250, 500, and 1000 µg/mL) inhibited the growth of MHCC97H cells in a dose-dependent manner via G2/M phase cell cycle arrest with no evidence of apoptosis. Among the identified proteins with upregulated expression were dynactin subunit 2, N-myc downstream-regulated gene 1, heat shock protein beta-1, alpha-enolase isoform 1, phosphatidylinositol transfer protein, and WD repeat-containing protein 1. Meanwhile, the proteins with downregulated expression were 14-3-3 gamma, BUB3, microtubule-associated protein RP/EB family member 1, thioredoxin-like protein, chloride intracellular channel protein 1, ectonucleoside triphosphate diphosphohydrolase 5, xaa-Pro dipeptidase, enoyl-CoA delta isomerase 1, protein-disulfide isomerase-related chaperone Erp29, hnRNP 2H9B, peroxiredoxin 1, WD-40 repeat protein, and serine/threonine kinase receptor-associated protein. CONCLUSION: The water extract of C. cicadae reduced the growth of human hepatocellular carcinoma MHCC97H cells via G2/M cell cycle arrest.

Microbubble-mediated sonoporation amplified lipid peroxidation of Jurkat cells.

Sonoporation is a developing technique used in drug delivery for cancer cells. Low frequency ultrasound is used to trigger the cavitation of microbubbles to puncture the cell membrane, and during this process, lipid metabolism becomes disrupted. In this study, cell viability and the generation of specific oxidized lipid products were assessed in Jurkat cells before and after sonoporation. A reduction in cell viability and an induction of apoptosis of Jurkat cells were found 4 h and 24 h post-sonoporation, respectively. Sonoporation suppressed cholesterol concentration and arachidonic, eicosapentaenoic and docosahexaenoic acids in the Jurkat cells. Levels of enzyme-independent oxidized products (F2-isoprostanes, F3-isoprostanes, 7-ketocholesterol) were elevated by sonoporation compared with the control, whereas enzyme-dependent oxidized products (5(S)-, 9(S)-, 12(S)-, 15(S)- and 20-HETE and 27-hydroxycholesterol) were not altered. Antioxidant enzymes activities were also increased in sonoporated Jurkat cells compared with the control. In this study, the loss of lipids potentially increased the availability for enzyme-independent lipid peroxidation, leading to cell fragility and death.

Profiling of oxidized lipid products of marine fish under acute oxidative stress.

Free radical products including reactive oxygen species are potent to oxidize lipids and reliable measurements have been established mostly in human and rodent. To date, robust biomarkers were not used to assess the peroxidation in marine fish. The changes of oxidized lipid products from polyunsaturated fatty acids and cholesterol were assessed after exposure of H(2)O(2) to fish (medaka). Oxidized lipid products released by free radical reaction (F(2)-isoprostanes and metabolites, F(3)-isoprostanes, neuroprostanes, 7-ketocholesterol, 7ß-hydroxycholesterol), by lipoxygenase enzymes (5(S)-, 8(S)-, 12(S)- and 15(S)-HETE, and resolvin D1) and by cytochrome P450 (9(S)-, 11(S)- and 20-HETE, and 27-hydroxycholestrol) were measured in fish muscle using LC/MS/MS. Arachidonate, docosahexaenoate, eicosapentaenoate and cholesterol levels, and antioxidant enzymes activity (catalase, SOD and gluthathione reductase) measurement were also determined. Activity of antioxidant enzymes especially catalase were elevated in presence of H(2)O(2) however longer exposure time suppressed the antioxidant activities. Arachidonate, docosahexaenoate, eicosapentaenoate and cholesterol levels were reduced in presence of H(2)O(2) and oxidized lipid products (isoprostanes, neuroprostanes 5(S)-HETE, 20-HETE, 7-ketocholesterol, 27-hydroxycholesterol and resolvin D1) were rapidly released in the fish muscle. This study validates oxidized lipid products, noticeably isoprostanes are measurable in marine fish muscle and should be considered when assessing oxidative stress especially due to exogenous factors.

Limited antioxidant effect after consumption of a single dose of tomato sauce by young males, despite a rise in plasma lycopene.

This study investigated the effect of a single dose of tomato sauce on healthy male volunteers in a randomized crossover study. Healthy male subjects (n = 10) were enrolled. Placebo (rice and olive oil) or tomato (tomato sauce, rice and olive oil) meals were provided to the volunteers. Blood and urine samples were taken before consumption of meal (0 h) and 2, 4, 6, 24 and 48 h after meal. Consumption of tomato sauce increased plasma lycopene level by 5-22%, with a maximum level at 24 h (p<0.01) after the meal. Levels of plasma F(2)-isoprostanes, hydroxyeicosatetraenoic acid products, allantoin and urinary 8-hydroxy-2'-deoxyguanosine did not change after either meal, but urinary F(2)-isoprostanes (p<0.05) significantly decreased at 48 h compared to 0 h after the tomato sauce meal. This study showed that a single dose of tomato sauce meal had only a limited antioxidant effect in vivo.

Short-term control of capsaicin on blood and oxidative stress of rats in vivo.

Capsaicin (8-methyl-n-vanillyl-6-nonenamide), a pungent component found in red pepper can induce body heat and possibly enhance blood flow as well as increase energy expenditure, and prevent oxidative stress. Male Wistar rats were divided into vehicle, 1 mg/kg body weight capsaicin and 3 mg/kg body weight capsaicin groups. Samples were taken from the animals on day 1 of i.p. treatment with capsaicin and on 3 consecutive days of i.p. treatment with capsaicin. Our investigation demonstrated that blood flow measurements in rats was negatively correlated with LDL after treatment with capsaicin. Although capsaicin did not show a noticeable effect on the serum total cholesterol level, LDL decreased while HDL and triglyceride increased in rats treated with 3 mg/kg capsaicin for 3 days. The antioxidant effect of capsaicin was not shown when the rats were treated with 1 mg/kg body weight capsaicin. However, rats treated with 3 mg/kg body weight capsaicin for 3 days showed a reduction of oxidative stress measured as malondialdehyde in the liver, lung, kidney and muscle. Liver glycogen was found to decrease after 3 days treatment with 3 mg/kg body weight capsaicin. From this study, it is hypothesized that capsaicin can be a potent antioxidant and aid in lowering LDL even when consumed for a short period.

Immunoregulatory and antioxidant performance of alpha-tocopherol and selenium on human lymphocytes.

The role of alpha-tocopherol (alpha-toco) and selenium (Se) on human lymphocyte oxidative stress and T-cells proliferation were studied by flow cytometry. We measured the hydrogen peroxide and glutathione levels in cultured human T-lymphocytes and the proliferation of their subsets: T-helper/inducer, T-suppressor/cytotoxic, and natural killer and interleukin-2 receptors upon stimulation by the mitogens phytohemaglutinin (PHA) and lipopolysaccharide (LPS). The results indicate that early stimulation by mitogens is affected by the glutathione and hydrogen peroxide status of the T-lymphocytes. The addition of 100 microM or 500 microM alpha-toco or 0.5 microM Se alone shows weak antioxidant and immunostimulant properties. When combined, an enhanced antioxidant and immunoregulatory effect was observed. The present findings indicate that alpha-toco and Se have interactive effects as oxygen radical scavengers, thus promoting human lymphocyte response to antigens. This suggests that micronutrient status is an important factor in considering when interpreting the results of in vitro assays of lymphocyte function.