ABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Pleurotus eryngii (DC. ex Fr.) Quel has been collected from the wild, cultivated and used in traditional medicines to treat various disorders and diseases since antiquity. In traditional Chinese medicine, the powdered fruiting bodies of Pleurotus eryngii were used for immunostimulation, skin-care, wound-healing, cancer and lumbago treatment. In the current study, we investigated the antiproliferative activity of Pleurotus eryngii powder on A549, BGC-823, HepG2 and HGC-27 cancer cells and its immunomodulating activity on macrophage, RAW 264.7 cells based on its active compound. MATERIALS AND METHODS: A novel bioactive protein (PEP) was extracted from Pleurotus eryngii fruiting bodies powder and purified on DEAE-52, CM-52 and Superdex 75 column chromatographies using an ÄKTA purifier. Its cytotoxicity on A549, BGC-823, HepG2, HGC-27 and RAW 267.4 cell lines was then evaluated using MTT, alamar blue (AB), trypan blue (TB), neutral red (NR), lactate dehydrogenase (LDH), Annexin V FITC/PI and morphological change assays. Moreover, lysosomal enzyme activity, pinocytosis, nitric oxide (NO) and hydrogen peroxide (H2O2) production assays were used to examine immunomostimulatory activity of PEP on RAW 267.4 cells. RESULTS: Based on high performance gel permeation chromatography (HPGPC), Fourier transform infrared (FT-IR) and nuclear magnetic resonance (NMR) analyses, the isolated protein (PEP) had a molecular weight of 63 kDa, a secondary (α-helical) structure and was mainly composed of arginine, serine and glycine. PEP significantly (P<0.05) inhibited A549, BGC-823, HepG2 and HGC-27 tumor cells proliferation dose-dependently with an IC50 range of 36.5 ± 0.84 to 229.0 ± 1.24 µg/ml. Contrarily, PEP stimulated the proliferation of macrophages. CONCLUSION: Pleurotus eryngii fruiting bodies powder has a potential application as a natural antitumor agent with immunomodulatory activity, proposedly, by targeting the lysosomes of cancerous cells and stimulating macrophage-mediated immune responses.
Subject(s)
Antineoplastic Agents/pharmacology , Immunologic Factors/pharmacology , Peptides/pharmacology , Pleurotus , Animals , Cell Line , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Complex Mixtures/chemistry , Fruiting Bodies, Fungal , Humans , Hydrogen Peroxide/metabolism , Lysosomes/metabolism , Macrophages/drug effects , Macrophages/physiology , Mice , Nitric Oxide/metabolism , Pinocytosis/drug effects , PowdersABSTRACT
BACKGROUND: Currently, the use of mushrooms as functional foods, nutraceuticals or phytopharmaceuticals source has risen. In contrast, the possible cellular cytotoxicity and immunostimulatory activity of Pleurotus eryngii (DC. ex Fr.) Quel protein (PEQP) is unknown. Here we report extraction, anti-tumorigenic and immunostimulatory activity of PEQP in vitro. RESULTS: PEQP was extracted from the dried fruiting bodies of P. eryngii, purified and characterised. Its in vitro antiproliferative activity was then evaluated in human non-small cell lung cancer A549 (NSCLC), stomach adenocarcinoma BGC-823, hepatocellular carcinoma HepG2 and gastric carcinoma HGC-27 cell lines using conventional cancer drugs (paclitaxel, doxorubicin and mitomycin C) as positive controls. The protein fractions (PEQP 1, 2, 3 and 4) obtained inhibited tumour cell proliferation dose-dependently with fraction PEQP 2 having significant (P < 0.05) toxicity in all tumour cells. PEQP had no significant toxicity on normal liver Chang cells but their proliferation was significantly inhibited by mitomycin C. Moreover, PEQP stimulated the proliferation, lysosomal enzyme activity, pinocytosis, nitric oxide and hydrogen peroxide production of RAW 264.7 cell lines dose-dependently. CONCLUSION: Based on these results, P. eryngii protein has a potential application in functional foods as a natural anti-tumour agent with immunostimulatory activity.
