Branched mannans from the mushroom Cantharellus cibarius enhance the anticancer activity of natural killer cells against human cancers of lung and colon.

Several studies have shown that mushroom polysaccharides enhance the ability of natural killer (NK) cells to recognize cancer cells as foreign and thereby enhance the effectiveness of host immune defence mechanisms. Nevertheless, the use of NK cells in cancer treatment requires finding selective stimulators of their cytotoxicity without disturbing organism homeostasis. Our studies revealed that Cantharellus cibarius polysaccharides present in the CC2a fraction, mainly composed of an O-2 and O-3 branched (1→6)-linked mannan, not only beneficially influenced the viability and proliferation of the human natural killer cells NK92 but also enhanced their anticancer properties against the human lung and colon cancer cells A549 and LS180, and at the same time did not affect the human lung and colon epithelial cells NL20 and CCD841 CoN. Furthermore, the CC2a fraction used alone was also nontoxic to the normal epithelium, while it inhibited the viability of these cancer cells. Nevertheless, the therapeutic potential of NK92 cells was greatly enhanced after coincubation with these polysaccharides and the observed effect was dependent on the CC2a concentrations. The beneficial effect of CC2a on NK92 cells was associated with stimulation of p38 and Erk expression as well as induction of the transcription factor CREB. The discovered beneficial impact of the CC2a fraction on NK92 cells suggested the therapeutic use of the investigated compound especially as an adjuvant. Furthermore, taking into account the abundance of these water soluble mannans in C. cibarius, the results also suggest that an increase in the intake of C. cibarius may promote innate immunity response against cancer through the enhancement of NK cell activity.

Cantharellus cibarius branched mannans inhibits colon cancer cells growth by interfering with signals transduction in NF-ĸB pathway.

In order to evaluate the therapeutic potential of polysaccharides obtained from the widely consumed Cantharellus cibarius mushroom on NF-ĸB pathway involved in cancer cells proliferation, survival, and metastasis, their antiproliferative and cytotoxic properties in human colon cancer cells LS180 and human colon epithelial cells CCD841 CoN were studied. BrdU and LDH assays results show that a branched mannan isolated from C. cibarius (CC2a) was selective against colon cancer cells, suppressing their proliferation and destroying membrane integrity but at the same time not adversely affecting colon epithelial cells. Results of Western blotting, immunofluorescence, RealTime PCR revealed that CC2a anticancer abilities were accompanied by disorders in signals transduction in NF-ĸB pathway in particular inhibition of IκBα degradation, attenuation of activated NF-κB phosphorylation and the subsequent decrease of NF-κB nuclear level as well as significant down-regulation of NF-κB target genes BAX, BCL2, CCND1, MMP9, MYC, BIRC5 and the corresponding proteins (except Bax). Furthermore, CC2a treatment of LS180 cells resulted in perturbation in G0/G and S phases of the cell cycle also associated with a marked increase of DNA fragmentation as well as inhibition of cancer cells motility. Obtained results indicated C. cibarius branched mannans as a new option in fighting with colon cancer.

Mushroom small RNAs as potential anticancer agents: a closer look at Cantharellus cibarius proapoptotic and antiproliferative effects in colon cancer cells.

Screening aimed at the evaluation of the presence of small RNAs with anticancer properties in three mushrooms species, besides Boletus edulis, namely Boletus spretus (current name Baorangia emilei), Boletus pinophilus and Cantharellus cibarius, was conducted. All mushrooms yielded an ethanol insoluble and water soluble small RNA fraction purified from co-extracted polysaccharides by anion-exchange chromatography. Small RNAs from B. spretus and C. cibarius showed strong antiproliferative activity against human colon adenocarcinoma cell lines (IC50 of 5.6 µg mL-1 and 11.1 µg mL-1 for LS180 and 1.9 µg mL-1 and 12.6 µg mL-1 for HT-29 cell lines, respectively) while those isolated from B. pinophilus showed a much lower antiproliferative activity in these cells. All RNA fractions were nontoxic against CCD841 CoTr human colon epithelial cells. A detailed study of the anticancer mechanism of C. cibarius small RNAs showed that their antiproliferative activity was due to p53-dependent cell cycle arrest mediated by p21, while the proapoptotic effect was mostly dependent on the enhancement of p53 expression. Overall, small RNA fractions isolated from some edible mushrooms, namely C. cibarius, show potent antiproliferative activity without cytotoxicity to normal cells, being a potential new anticancer agent naturally present in mushrooms that we eat.

Evaluation of the effect of 2-(2,4-dihydroxyphenyl)-4H-benzofuro[3,2-d][1,3]thiazin-4-one on colon cells and its anticancer potential.

In this paper, we present the biological effect of the newly synthesized 2-(2,4-dihydroxyphenyl)-4H-benzofuro[3,2-d][1,3]thiazin-4-one (DPBT) on human colon adenocarcinoma cell lines (HT-29 and LS180). Additionally, DPBT cytotoxicity was examined in human colon epithelial cells (CCD 841 CoTr) and human skin fibroblasts (HSF). The studies revealed a significant decrease in the proliferation of cancer cells after exposure to DPBT at concentrations in the range of 10-100 µM. Additionally, DPBT was not toxic to normal CCD 841 CoTr and HSF cells at concentrations that induced inhibition of cancer cell proliferation. The nature of the anti-proliferative action of DPBT in the cell cycle progression in colon cancer cells and the expression of proteins involved in this process were examined by flow cytometry and western blotting, respectively. The investigations demonstrated higher sensitivity of LS180 than HT-29 to the DPBT treatment. The anti-proliferative action of DPBT in LS180 was attributed to cell cycle arrest in the G1 phase via up-regulation of p27KIP1 and down-regulation of cyclin D1 and CDK4 proteins.

Neuroprotective properties of Cantharellus cibarius polysaccharide fractions in different in vitro models of neurodegeneration.

The percentage of people suffering from neurodegenerative diseases is constantly increasing, because of that searching for substances able to prevent or inhibit neuronal death sseems to be reasonable. Because of the high popularity the search of new neuroprotective agents we started from Cantharellus cibarius. Neuroprotective properties of C. cibarius polysaccharides fractions was investigated in different models of neurodegeneration including trophic stress, excitotoxicity and andoxidative stress. Fractions influence on neurons viability was examined using Neurite Outgrowth Staining, MTT and LDH tests, while antioxidant capacity was determined by commercial antioxidant assays. Performed studies revealed beneficial effect of C. cibarius fractions (CC2a, CC3) on neurons viability and neurite outgrowth in normal and different stress conditions. Both tested fractions have shown antioxidant capacity and effectively neutralize the negative changes induced by glutamatergic system activators. Discovered neuroprotective properties of investigated compounds suggested the their use for developing effective and safety therapeutic strategy for neurodegenerative diseases.

Anticancer effect of the water extract of a commercial Spirulina (Arthrospira platensis) product on the human lung cancer A549 cell line.

Spirulina is a well-described and popular dietary supplement derived from Arthrospira algae. In the present study, the anticancer potential of a water extract of a commercial Spirulina product (SE) against the human non-small-cell lung carcinoma A549 cell line was evaluated. After qualitative analysis, we investigated the effect of SE on cell viability, proliferation, and morphology. Furthermore, the influence of SE on regulation of the cell cycle, induction of apoptosis in lung cancer cells, and expression of cell cycle/apoptosis-related proteins was evaluated. Additionally, we examined the cytotoxic effect of SE on normal human skin fibroblasts (HSF). Our studies revealed that SE significantly reduced cancer cell viability and proliferation, which was accompanied by cell cycle inhibition in the G1 phase, induction of apoptosis, and prominent morphological changes. Moreover, we detected no cytotoxic effect of the tested Spirulina extract on normal skin fibroblasts. Our molecular studies demonstrated that SE reduced the phosphorylation of Akt and Rb proteins, reduced the expression of cyclin D1 and CDK4, and increased the Bax to Bcl-2 ratio in the A549 cells. In conclusion, the results obtained provide evidence of the anti-cancer activity of the commercial Spirulina product against lung cancer cells and strongly support the knowledge of the chemopreventive properties of Spirulina.

Fomitopsis betulina (formerly Piptoporus betulinus): the Iceman's polypore fungus with modern biotechnological potential.

Higher Basidiomycota have been used in natural medicine throughout the world for centuries. One of such fungi is Fomitopsis betulina (formerly Piptoporus betulinus), which causes brown rot of birch wood. Annual white to brownish fruiting bodies of the species can be found on trees in the northern hemisphere but F. betulina can also be cultured as a mycelium and fruiting body. The fungus has a long tradition of being applied in folk medicine as an antimicrobial, anticancer, and anti-inflammatory agent. Probably due to the curative properties, pieces of its fruiting body were carried by Ötzi the Iceman. Modern research confirms the health-promoting benefits of F. betulina. Pharmacological studies have provided evidence supporting the antibacterial, anti-parasitic, antiviral, anti-inflammatory, anticancer, neuroprotective, and immunomodulating activities of F. betulina preparations. Biologically active compounds such as triterpenoids have been isolated. The mushroom is also a reservoir of valuable enzymes and other substances such as cell wall (1→3)-α-D-glucan which can be used for induction of microbial enzymes degrading cariogenic dental biofilm. In conclusion, F. betulina can be considered as a promising source for the development of new products for healthcare and other biotechnological uses.

Kynurenic Acid Induces Impairment of Oligodendrocyte Viability: On the Role of Glutamatergic Mechanisms.

Kynurenic acid (KYNA) is an end stage product of tryptophan metabolism with a variety of functions in the human body, both in the central nervous system (CNS) and in other organs. Although its activity in the human brain has been widely studied and effects on neural cells were emphasized, the effect of KYNA on oligodendroglial cells remains unknown. Present study aims at describing the activity of high concentration of KYNA in OLN-93 cells. The inhibition of OLN-93 oligodendrocytes viability by KYNA in a medium with reduced serum concentration has been demonstrated. Although decreased metabolic activity of KYNA treated OLN-93 cells was shown, the cells proliferation was not altered. KYNA treatment did not alter morphology as well as expression level of cell cycle and proliferation regulating proteins. Furthermore, glutamate receptor antagonists and agonists did not alter the inhibitory effect of KYNA on viability of OLN-93 oligodendrocytes. This study contributes to the elucidation of effects of KYNA on oligodendrocytes in vitro, yet further analyses are necessary to explain the mechanisms behind the damage and loss of myelin sheaths.

Cultivation and utility of Piptoporus betulinus fruiting bodies as a source of anticancer agents.

Piptoporus betulinus is a wood-rotting basidiomycete used in medicine and biotechnology. However, to date, no indoor method for cultivation of this mushroom fruiting bodies has been developed. Here we present the first report of successful production of P. betulinus mature fruiting bodies in artificial conditions. Four P. betulinus strains were isolated from natural habitats and their mycelia were inoculated into birch sawdust substrate supplemented with organic additives. All the strains effectively colonized the medium but only one of them produced fruiting bodies. Moisture and organic supplementation of the substrate significantly determined the fruiting process. The biological efficiency of the P. betulinus PB01 strain cultivated on optimal substrate (moisture and organic substance content of 55 and 65 and 25 or 35 %, respectively) ranged from 12 to 16 %. The mature fruiting bodies reached weight in the range from 50 to 120 g. Anticancer properties of water and ethanol extracts isolated from both cultured and nature-derived fruiting bodies of P. betulinus were examined in human colon adenocarcinoma, human lung carcinoma and human breast cancer cell lines. The studies revealed antiproliferative and antimigrative properties of all the investigated extracts. Nevertheless the most pronounced effects demonstrated the ethanol extracts, obtained from fruiting bodies of cultured P. betulinus. Summarizing, our studies proved that P. betulinus can be induced to fruit in indoor artificial culture and the cultured fruiting bodies can be used as a source of potential anticancer agents. In this respect, they are at least as valuable as those sourced from nature.

New derivative of 2-(2,4-dihydroxyphenyl)thieno-1,3-thiazin-4-one (BChTT) elicits antiproliferative effect via p38-mediated cell cycle arrest in cancer cells.

2-(2,4-Dihydroxyphenyl)thieno-1,3-thiazin-4-ones are a group of new compounds with potential anticancer activity. This type of derivatives was poorly investigated in the area of synthesis and biological activities. In the present study the antiproliferative action of the most active derivative BChTT was described. The aim of biological evaluation was to investigate the ability of the compound to inhibit cancer cell proliferation and identify mechanism involved in its action on the molecular level. BChTT inhibited the proliferation of lung cancer A549, colon cancer HT-29 and glioma C6 cells in the concentration-dependent manner. It was not toxic to normal cells including skin fibroblasts, hepatocytes and oligodendrocytes in the antiproliferative concentrations. BChTT decreased the DNA synthesis in the treated cancer cells and induced cell cycle arrest in the G0/G1 phase. Moreover, the ability of the compound to activate p38 kinase and decrease cyclin D1 expression was estimated. Participation of p38 kinase in the antiproliferative action of the compound was confirmed by the analysis of BChTT activity in the cells with the p38 silenced gene. The obtained results may suggest the ability of the tested derivative to inhibit cancer cells proliferation by induction of p38-mediated cyclin D1 downregulation.

Synthesis of 2-(2,4-dihydroxyphenyl)thieno-1,3-thiazin-4-ones, their lipophilicity and anticancer activity in vitro.

A new one-step synthesis of novel biologically active 2-substituted 2,4-dihydroxyphenyl-4[Formula: see text]-thieno[3,2-[Formula: see text]][1,3]thiazin-4-ones and 4[Formula: see text]-thieno[2,3-[Formula: see text]][1,3]thiazin-4-ones has been elaborated and described. The compounds were prepared by the reaction of aryl-modified sulfinylbis [(2,4-dihydroxyphenyl)methanethione]s and the corresponding aminothiophenecarboxamides. The derivatives showed anticancer activity in vitro. These compounds inhibited the proliferation and viability of lung cancer A549, colon cancer HT-29 and glioma C6 cells in a concentration-dependent manner. Some of the derivatives had no influence on normal skin fibroblasts culture viability. Moreover, one compound (1b) showed the ability to inhibit DNA synthesis in cancer cells, especially in C6 cells, and was not toxic for normal oligodendrocytes and hepatocytes. Using reversed phase RP 18 HPLC and immobilised artificial membrane (IAM) chromatography the phase affinity of the compounds was determined. The influence of lipophilicity on the activity of compounds has been discussed.