Polysaccharide Extracted From Pleurotus Ostreatus Possesses Antitumor Properties Against Hepatocellular Carcinoma And H22 Mouse Hepatoma Ascites Model In Vivo.

Background: In Homo sapiens,any diseased condition related to abnormal growth of cells is referred to as Cancer.Globally every year the rate of patients suffering from this non-communicable disease is rising.Hepatocellular carcinoma is one of the leading causes of mortality and morbidity among human beings.Identification and treatment progress of hepatocarcinoma is not obvious to understand.Many new biomarkers need to be discovered for the exact diagnosis and treatment of HCC.Many new cases reported of HCC showed resistance towards most of the chemotherapeutic drugs.That makes the scientist thinking about alternative medicine therapy or supportive drug therapy for the patients suffering from HCC.The ancient history of mankind shows that natural products derived from plants,fungi,and micro-organisms used for culinary purposes and some of them bearing exceptional therapeutic attributes and over the period show some evidence that compounds and byproducts derived from these compounds can be used for a purpose of therapeutics.Mushrooms having therapeutic potential are the main focus these days for treating cancer patients either as a supportive drug system or anti-tumor drug.Proteins,carbohydrates,protein-protein complex,carbohydrate-protein complex,and other compositional elements can be used for their therapeutic capabilities.Pleurotus ostreatus(Pleurotus spp)is one of the most edible cuisines in Asia as well as all over the world,it has been a source of nutritious diet since it was classified as an edible mushroom with no or negligible side effects.Pleurotus species were reported in more than several cases for their therapeutic potentials and activities against cancer.Pleurotus ostreatus is one of the famous mushrooms for its therapeutically uses and most significantly it was reported to possess anti-cancerous activity.Malignant ascites are a highly severe and intractable complication of advanced or recurrent malignant tumors that is often immunotherapy-resistant.Pleurotus ostreatus is widely used in traditional medicine as an anti-microbial and anti-cancer agent,but its effectiveness in treating malignant ascites is unclear.Objectives 1.Extraction of polysaccharides from the Pleurotus ostreatus and analyze the composition of monosaccharide and their molecular weight.Purify the polysaccharide and used it against hepatocellular carcinoma and h22 hepatoma in the mice model.2.In vivo study for the anti-tumor and immunological modulation of extracted polysaccharide used as a drug in malignant ascites mice model.3.In vitro analysis of the anti-tumor activity of extracted polysaccharide against hepatocellular carcinoma cell lines.Methods: Polysaccharides extracted from Pleurotus ostreatus by Phenol-sulphuric acid method,and then further classified the monosaccharide composition of polysaccharides extracted from Pleurotus ostreatus.H22 malignant ascites model was developed in Balb/C mouse for observing the effect of anti-tumor capabilities of polysaccharide in vivo,whereas Elisa and Immunohistochemistry were performed to further investigate the effects of polysaccharide in vivo.To observe the polysaccharide anti-tumor property of polysaccharides in HCC cell lines derived from homosapiens origin we used proliferation assays such as cell viability assay and Colonogenic assay to observe the colony formation capabilities of cancer cells.For further investigation of anti-cancer activities of Pleurotus ostreatus polysaccharide,we performed relative gene expression by quantitative PCR and western blot method.A Transwell chamber assay was performed to observe the migration and invasion properties of HCC cell lines.To observe the metastatic properties of HCC cell lines we perform a wound-healing assay.Immunofluorescence assay was performed to observe the gene expression of Cytochrome C supposed to be the initiator of apoptosis.Results: Polysaccharide extracted from Pleurotus ostreatus significantly regulates the immune response in the H22 malignant ascites model in mice and genetic expression of genes responsible for the regeneration of disease decreased as the dose increased.As well as the effects of polysaccharides in vitro polysaccharides showed anti-proliferative,anti-metastatic,and anti-migratory capability.Relative gene expression of genes involved in apoptosis was observed that shows the polysaccharide capable of inducing apoptosis in HCC cell lines by targeting the major genes involved in apoptosis generally.Conclusion: Conclusive results of our research findings conclude that the polysaccharide extracted from the Pleurotus ostreatus has the potential to be used as an anti-tumor drug.Comprehensive pieces of evidence proved that polysaccharide has the potential to be served as an antiproliferative agent against HCC cell line and also induce an immune response in mice model.It can also help decrease the migration invasion and metastatic properties of cancer cells.Those results show tremendous potential in this polysaccharide to be used against cancer.