Study On Activity And Mechanism Of Active Compounds Of Taiwanofungus Camphoratus Against Non-small Cell Lung Cancer

Aim: Lung cancer is one of the world’s highest malignancies.Non-small cell lung cancer accounts for about 80% of all types of lung cancer,its prevention,diagnosis,treatment and mechanism of research has become the focus of attention of many pharmaceutical scholars.At present,the main treatment of non-small cell lung cancer is chemotherapy,however,the current clinical chemotherapy drugs generally have a profound side effect,and easy to produce drug resistance.Medicinal fungi have a unique economic and pharmaceutical value in the East.The study of the active component from natural products has attracted many researchers.Therefore,to extract high efficiency and low toxicity natural anti-tumor components from medicinal fungus,can develop the new drug market,and will be the hot and difficult parts of the study to pharmaceutical scholars.Taiwanofungus camphoratus is a rare medicinal fungus native to Taiwan,which has a variety of physiological activities,especially the excellent anti-tumor activity.Antroquinonol,a star molecule from Taiwanofungus camphoratus,has an excellent inhibitory effect on non-small cell lung cancer,and now entered the FDA approved non-small cell lung cancer Phase II clinical trials.However,Antroquinonol mainly derived from the solid wood cultivated fruiting body,due to the camphor tree is a rare protection tree species,and was deforested,the research cost is very high,furthermore,the growth of solid fruiting body is extremely slow,it is not conducive to scientific research.Liquid fermentation has numerous advantages of a wide range of raw materials,low cost,short growth cycle and easy operation,to separate and purify the active component from the liquid fermentation products,is a very worthy way to develop.Method: In this study,crude extract of Taiwanofungus camphoratus liquid fermentation was first to use as the material to study its inhibitory effect on non-small cell lung cancer.It exhibited inhibitory effect on non-small cell lung cancer and caused S-cell cycle arrest,induced cell apoptosis.However,the crude extract is a mixture of many active components,and it is difficult to find out the real active monomeric component.Therefore,the follow-up study using the two monomeric compounds isolated from the laboratory earlier.By-1 was mainly from the fermentation broth.Antrodin C was mainly from the mycelium grown in submerged culture.The aim is to assess the inhibitory for By-1 and Antrodin C for NSCLC,and further explore its mechanism.The anti-lung cancer activity of By-1 and Antrodin C were investigated by detecting cell migration,cell proliferation,cell cycle,apoptosis and apoptosis-related proteins and autophagy,and further to explore whether the independent regulation of autophagy and apoptosis involved in lung cancer activity,as well as the signaling pathways.Result: It was found that both By-1 and Antrodin C exhibited an inhibitory effect on non-small cell lung cancer SPCA-1 cell line and inhibited the migration of SPCA-1 cells as well.The mechanism of inhibition of NSCLC is due to the S-cell cycle arrest,up-regulation of P53 protein,and P53 involved in ROS and Bcl-2-mediated mitochondrial cell apoptosis pathway,and further cause cascade of Caspase family to promote apoptosis factor Caspase-3 release,thus induced apoptosis to show the inhibition of SPCA-1.The autophagic structure was found by detection of autophagosome,autophagic flux and autophagy marker protein LC3 II.It was confirmed that By-1 and Antrodin C induced autophagy in SPCA-1 cells.And further found that autophagy was down-regulated through Akt / mTOR pathway.Through treatment of autophagic inhibitor and autophagic activator,the cell proliferation and apoptosis was detected,autophagy induced by By-1 and Antrodin C play a protective role in anti-lung cancer activity,and the co-treatment with pro-apoptotic agent and anti-autophagy agents may be a useful strategy in enhancingthe anticancer efficacy in NSCLC.There are numerous types of gene mutations in lung cancer cells,where EGFR mutations account for more than half of the total mutation type.Studies have shown that the use of targeted therapy method to treat the EGFR mutant lung cancer has higher efficiency,and the prognosis is good.The detection of cell proliferation,cell cycle and apoptosis in EGFR mutant cell line HCC827 showed that By-1 and Antrodin C inhibited the cell proliferation,caused the S phase cell cycle arrest and induced cell apoptosis.In this study,different NSCLC cell lines were used in a study of the anti-lung cancer and its molecular mechanism of By-1 and Antrodin C.By-1 and Antrodin C exhibited an inhibitory effect on NSCLC,the mechanism is regulated by the interaction between apoptosis and autophagy.Thus,By-1 and Antrodin C are likely to develop into potential anti-lung cancer treatment of chemotherapy drugs,which also provide theoretical basis in-depth study.