epiglucan and Liver-Neoplasms

Maitake mushroom (Grifola frondosa) MD-fraction containing beta-1,6 glucan with beta-1,3 branched chains has previously exhibited strong anticancer activity by increasing immune-competent cell activity.1,2 In this non-random case series, a combination of MD-fraction and whole maitake powder was investigated to determine its effectiveness for 22- to 57-year-old cancer patients in stages II-IV. Cancer regression or significant symptom improvement was observed in 58.3 percent of liver cancer patients, 68.8 percent of breast cancer patients, and 62.5 percent of lung cancer patients. The trial found a less than 10-20 percent improvement for leukemia, stomach cancer, and brain cancer patients. Furthermore, when maitake was taken in addition to chemotherapy, immune-competent cell activities were enhanced 1.2-1.4 times, compared with chemotherapy alone. Animal studies have supported the use of maitake MD-fraction for cancer.

Topics: Adult; Agaricales; Animals; Antibiotics, Antineoplastic; beta-Glucans; Breast Neoplasms; Carcinoma; Carcinoma, Hepatocellular; Carcinoma, Intraductal, Noninfiltrating; Carcinoma, Squamous Cell; Dose-Response Relationship, Drug; Female; Glucans; Humans; Liver Neoplasms; Lung Neoplasms; Male; Middle Aged; Neoplasms

Although macrophages contribute to cancer cell dissemination, immune evasion, and metastatic outgrowth, they have also been reported to coordinate tumor-specific immune responses. We therefore hypothesized that macrophage polarization could be modulated therapeutically to prevent metastasis. Here, we show that macrophages respond to β-glucan (odetiglucan) treatment by inhibiting liver metastasis. β-glucan activated liver-resident macrophages (Kupffer cells), suppressed cancer cell proliferation, and invoked productive T cell-mediated responses against liver metastasis in pancreatic cancer mouse models. Although excluded from metastatic lesions, Kupffer cells were critical for the anti-metastatic activity of β-glucan, which also required T cells. Furthermore, β-glucan drove T cell activation and macrophage re-polarization in liver metastases in mice and humans and sensitized metastatic lesions to anti-PD1 therapy. These findings demonstrate the significance of macrophage function in metastasis and identify Kupffer cells as a potential therapeutic target against pancreatic cancer metastasis to the liver.

Topics: Animals; beta-Glucans; Carcinoma, Pancreatic Ductal; Humans; Kupffer Cells; Liver Neoplasms; Mice; Pancreatic Neoplasms

Natural polysaccharide as the third abundant biomacromolecule has attracted considerable attentions due to their superior anti-tumor activities. However, the anti-tumor mechanism of polysaccharides has not been completely understood. Herein, the anti-tumor effects of black fungus polysaccharide (BFP), a typical β-glucan was comprehensively investigated, and the anti-tumor mechanism was obtained from metabolomics profiling. The in vitro results demonstrate that BFP inhibited the proliferation, migration and invasion of hepatoma carcinoma cells (HCC) through inducing the cell apoptosis and arresting the cell cycle at S phase without direct cytotoxicity. The hepatoma-bearing nude mice experiments further demonstrate that BFP could significantly inhibit the growth without system toxicity in vivo. Mass spectrometry-based metabolomics unveils that BFP significantly disturbed the multiple metabolic pathways, leading to the inhibition of tumor cells proliferation by promoting DNA damage, attenuating DNA damage repair, and inhibiting DNA synthesis. This study provides new insights for pharmacological research and clinical practice of polysaccharides.

Topics: Animals; Antineoplastic Agents; Apoptosis; Basidiomycota; beta-Glucans; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Proliferation; DNA Damage; DNA Repair; Fungal Polysaccharides; Humans; Liver Neoplasms; Male; Metabolome; Metabolomics; Mice, Inbred BALB C; Mice, Nude; S Phase Cell Cycle Checkpoints

The unique conformation transition from a triple helix to single coils for the triple helical β-d-glucans has paved the way to fabricate various functional nanocomposites through the denaturing-renaturing process. This study firstly reports a novel kind of naturally derived supramolecular polymer micelle consisting of single-stranded chains of curdlan (CUR) and β-CDs. It is proposed that β-CDs as the host molecules were threaded onto single β-glucan chains (denatured triplex CUR) via the host-guest interaction, thereby forming supramolecular micelles. The results from the 1H NMR, FT-IR, XRD and 2D 1H NOESY NMR studies confirmed the formation of the inclusion complex and the existence of the core-shell structure of the supramolecular assembly. TEM images and DLS revealed that the self-organized micelles displayed a regular spherical shape with an average diameter of ∼27 nm. Furthermore, the hydrophobic anticancer drug camptothecin (CPT) was selected as a model drug and successfully encapsulated into the CUR/β-CD micelles. The drug-loaded micelles exhibited a steady sustained-release pattern regardless of the environmental pH. The flow cytometry and confocal laser scanning microscopy measurements confirmed that the CPT-loaded micelles could be well internalized into HepG 2 cells and continuously release the drug molecules inside the tumor cells. Meanwhile, the in vivo experiments demonstrated that CPT-loaded micelles could effectively inhibit tumor growth in comparison to free drugs. This concept will give a favorable platform to construct intelligent drug delivery systems for potential use.

Topics: Animals; Antineoplastic Agents, Phytogenic; beta-Glucans; Camptothecin; Cell Proliferation; Cell Survival; Delayed-Action Preparations; Drug Compounding; Drug Delivery Systems; Hep G2 Cells; Humans; Liver Neoplasms; Mice; Micelles; Microscopy, Confocal; Models, Molecular; Protein Conformation, alpha-Helical; Spectroscopy, Fourier Transform Infrared; Xenograft Model Antitumor Assays

Ajwa dates (Phoenix dactylifera L.) are used by traditional therapeutic practitioners for several health benefits but most remain to be scientifically validated. In this study, we evaluated the apoptosis-inducing effect of ethanolic extract of Ajwa date pulp (ADP) on human hepatocellular carcinoma (HCC) HepG2 cells. High performance liquid chromatography analysis revealed the presence of polysaccharide β-D-glucan in ADP extract. Treated HCC cells revealed morphological characteristics of apoptosis under phase contrast microscopy. MTT assay demonstrated significant (p < 0.05) dose- and time-dependent inhibition of HCC cell growth. HCC cells were found to be in late apoptotic stage on treatment with higher doses of ADP extract as depicted by acridine orange/ethidium bromide and Annexin V-FITC/PI double stain. Importantly, ADP extract increased the reactive oxygen species level and decreased the mitochondrial membrane potential in treated HCC cells. Flow cytometry analysis demonstrated that ADP extract induced elevation of S and G2/M phases of cell cycle. Moreover, ADP extract induced apoptosis in HCC cells independent of tumor suppressor genes viz. CHEK2, ATM and TP53. Interestingly, ADP extract did not display any significant effect on normal cell line Vero. This study provides validation that ADP extract can be considered as a safe and natural potential drug candidate against human liver cancer.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; beta-Glucans; Carcinoma, Hepatocellular; Cell Cycle Checkpoints; Chlorocebus aethiops; Cytostatic Agents; Hep G2 Cells; Humans; Liver Neoplasms; Membrane Potential, Mitochondrial; Phoeniceae; Plant Extracts; Proteoglycans; Reactive Oxygen Species; Vero Cells

β-glucans are one of the most abundant forms of polysaccharides known as biological response modifiers which influence host's biological response and stimulate immune system. Accordingly, this study was initiated to evaluate irradiated β-glucan as a modulator for cellular signaling growth factors involved in the pathogenesis of hepatocellular carcinoma in rats. Hepatocellular carcinoma was induced with 20 mg diethylnitrosamine/kg BW. Rats received daily by gastric gavage 65 mg irradiated β-glucan/kg BW. It was found that treatment of rats with diethylnitrosamine induced hepatic injury and caused significant increase in liver injury markers with a concomitant significant increase in both hepatic oxidative and inflammatory indices: alpha-fetoprotein, interferon gamma, and interleukin 6 in comparison with normal and irradiated β-glucan-treated groups. Western immunoblotting showed a significant increase in the signaling growth factors: extracellular signal-regulated kinase 1 and phosphoinositide 3-kinase proteins in a diethylnitrosamine-treated group while both preventive and therapeutic irradiated β-glucan treatments recorded significant improvement versus diethylnitrosamine group via the modulation of growth factors that encounters hepatic toxicity. The transcript levels of vascular endothelial growth factor A and inducible nitric oxide synthase genes were significantly higher in the diethylnitrosamine-treated group in comparison with controls. Preventive and therapeutic treatments with irradiated β-glucan demonstrated that the transcript level of these genes was significantly decreased which demonstrates the protective effect of β-glucan. Histological investigations revealed that diethylnitrosamine treatment affects the hepatic architecture throughout the significant severe appearance of inflammatory cell infiltration in the portal area and congestion in the portal vein in association with severe degeneration and dysplasia in hepatocytes all over hepatic parenchyma. The severity of hepatic architecture changes was significantly decreased with both β-glucan therapeutic and preventive treatments. In conclusion, irradiated β-glucan modulated signal growth factors, vascular endothelial growth factor A, extracellular signal-regulated kinase 1, and phosphatidylinositol-3-kinase, which contributed to experimental hepatocarcinogenesis.

Topics: Animals; beta-Glucans; Carcinoma, Hepatocellular; Diethylnitrosamine; Disease Models, Animal; Gamma Rays; Humans; Liver; Liver Neoplasms; Mitogen-Activated Protein Kinase 3; Phosphatidylinositol 3-Kinases; Rats; Signal Transduction; Vascular Endothelial Growth Factor A

Two sulfated derivatives (PRP-S1 and PRP-S2) of a β-glucan from Phellinus ribis with different degrees of substitution were obtained by chlorosulfonic acid method. The derivatives could block formation of new vessels in zebrafish and inhibit the proliferation of human umbilical vein endothelial cells (HUVECs). The two sulfated derivatives had remarkably high antitumor activities in vivo (in BALB/c mice inoculated with H22 hepatocellular carcinoma) as well as in vitro (against human ovary cancer SKOV-3 cells), without producing any overt signs of general toxicity. The results of immunohistochemistry assay indicated that the derivatives significantly reduced the average number of microvessel density (MVD) and inhibited the expression of vascular endothelial growth factor (VEGF) in tumor. Thus, these derivatives exhibit pronounced antiangiogenic and antitumoral properties. Except for cytotoxic effects on tumor cells, it is reasonable to expect that the antitumoral effects of PRP-S1 and PRP-S2 are mediated via their antiangiogenic properties.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Basidiomycota; beta-Glucans; Carbohydrate Sequence; Carcinoma, Hepatocellular; Cell Proliferation; Cells, Cultured; Female; Glucans; Human Umbilical Vein Endothelial Cells; Humans; Immunoenzyme Techniques; Liver Neoplasms; Mice; Mice, Inbred BALB C; Molecular Sequence Data; Neovascularization, Pathologic; Ovarian Neoplasms; Xenograft Model Antitumor Assays; Zebrafish

Common genetic alteration in cancer genomes is implicated for embracing an aberrant cancer gene participated in tumor progression. In this study, we identified a somatic mutated LIM and cysteine-rich domains-1 (LMCD1) as a putative metastatic oncogene in human hepatocellular carcinoma (HCC) using integrated genomic approaches. In addition to revealing genomic amplification and gene upregulation, we identified recurrent E135K (3/48 cases) mutations in HCC tissues and K237R mutation in the PLC/PRF/5 HCC cell line. Expression of mutant LMCD1 E135K or K237R reduced the stress fiber assembly, increased cortical actin accumulation and induced lamellipodial extension. Consistently, these mutations enhanced cell migration and showed activation of the Rac1-signaling pathway. Inhibition of the LMCD1/Rac1 pathway by an LMCD1 short-hairpin RNA (shLMCD1) or the Rac1 inhibitor NSC23766 suppressed the mutation-mediated lamellipodial protrusion and cell migration. In PLC/PRF/5 cells with endogenous K237R mutation, cell migration was enhanced by estrogen-induced LMCD1 expression but reversed by shLMCD1 treatment. Moreover, overexpression of LMCD1 E135K mutation significantly promoted systemic lung metastasis in a murine tail vein injection model. Together, our results suggest that LMCD1 mutations are potential oncogenic events in HCC metastasis to promote cell migration through the Rac1-signaling pathway.

Topics: Animals; beta-Glucans; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Movement; Co-Repressor Proteins; Gene Amplification; Gene Knockdown Techniques; Humans; LIM Domain Proteins; Liver Neoplasms; Lung Neoplasms; Mice; Neoplasm Metastasis; Neoplasm Transplantation; Point Mutation; Pseudopodia; rac1 GTP-Binding Protein; Signal Transduction; Up-Regulation

The mushroom Agaricus blazei is studied for its nutraceutical potential and as a medicinal supplement. The aim of the present study was to investigate the chemoprotective effect of beta-glucan extracted from the mushroom A. blazei against DNA damage induced by benzo[a]pyrene (B[a]P), using the comet assay (genotoxicity) and micronucleus assay with cytokinesis block (mutagenicity) in a human hepatoma cell line (HepG2). To elucidate the possible beta-glucan mechanism of action, desmutagenesis or bioantimutagenesis types, three treatment protocols were tested: simultaneous, pre-treatment, and presimultaneous. The results showed that beta-glucan does not exert genotoxic or mutagenic effect, but that it does protect against DNA damage caused by B[a]P in every protocol tested. The data suggest that beta-glucan acts through binding to B[a]P or the capture of free radicals produced during its activation. On the other hand, the pre-treatment results also suggest the possibility that beta-glucan modulates cell metabolism.

Topics: Agaricus; Antimutagenic Agents; Benzo(a)pyrene; beta-Glucans; Carcinoma, Hepatocellular; Cell Line, Tumor; Comet Assay; DNA Damage; Drug Administration Schedule; Free Radical Scavengers; Free Radicals; Humans; Liver Neoplasms; Micronucleus Tests; Mutagens

The tumoricidal activity of a bioactive metabolite produced by submerged culture in a 2.1-L airlift fermentor of Grifola frondosa NTUS was investigated. After 14 days of cultivation, ethyl acetate extracts from the supernatant of culture broth (EES) were analyzed by cell viability assay. The IC50 of EES for cytotoxicity against human carcinoma cells (Hep 3B, Hep G2, HeLa, CL1-1) and normal human lung fibroblast MRC-5 was 78.4, 52.7, 77.6, 71.0, and 233.3 microg/mL, respectively. EES was further fractionated and a main cytotoxic compound, HE-5-5, was obtained. The IC50 of HE-5-5 based on the cell viability of Hep 3B and MRC-5 cells was 3.6 and 33.1 microg/mL, respectively. Thus, HE-5-5 showed a selective cytotoxic effect against Hep 3B cells and MRC-5. According to the UV, MS, and NMR data, HE-5-5 was identified as o-orsellinaldehyde. A DNA fragmentation assay together with the presence of a significant sub-G1 peak by flow cytometry suggested that o-orsellinaldehyde might mediate its cytotoxicity through apoptosis.

Topics: Aldehydes; Antineoplastic Agents; Apoptosis; beta-Glucans; Carcinoma, Hepatocellular; Catechols; Cell Cycle; Cell Line, Tumor; DNA Fragmentation; Grifola; Humans; Liver Neoplasms

The purpose of this study was to investigate the combined antitumor effect of aminated beta-1,3-D-glucan (AG) and interferon-gamma (IFN-gamma) in an experimental liver metastasis model. Liver metastases were established by inoculation of C-26 colon carcinoma cells into the superior mesenteric vein of syngeneic mice. Treatment of mice started 24 hours after inoculation of tumor cells by daily intravenous injections of either AG, IFN-gamma, or a combination of both for a duration of 6 days. The resultant liver metastases were then quantified after an additional period of 11 days. Combination of IFN-gamma and AG inhibited the growth of liver metastases almost entirely. IFN-gamma was also very efficient, while AG alone did not exert any significant antitumor effect. These results, along with histological studies from mice receiving AG and IFN-gamma, indicated that activation and recruitment of liver macrophages may be a part of the mechanism responsible for the inhibition of metastatic growth observed in this study.

Topics: Animals; Antineoplastic Agents; beta-Glucans; Carcinoma; Colonic Neoplasms; Drug Synergism; Female; Glucans; Interferon-gamma; Liver Neoplasms; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Neoplasm Metastasis; Recombinant Proteins