The Inhibitory Effect Of Water Extract Of Inonotus Obliquus On Human Hepatocellular Carcinoma Cell Line HepG2

Inonotus obliquus,also known as Chaga,is a medicinal fungus growing on birch.It contains a variety of chemical components and has high medicinal value.A large number of studies have shown that Inonotus obliquus has many functions,such as anti-oxidation,hypoglycemic,immunomodulatory and anti-inflammatory,but there are few studies on the anti-tumor effect and its mechanism of the crude extracts of Inonotus obliquus.This study will discuss the inhibitory effect of Inonotus obliquus water extract on the human hepatoma cell line HepG2.In this study,the effect of those six different extracts which are form Canadian and Russian Inonotus obliquus and 5-FU on antiproliferation of the human hepatocarcinoma cells HepG2,the human hepatocarcinoma cells Hep3B and the normal liver cells L02,was investigated by MTT method.The morphological changes on HepG2 were observed by the ordinary light microscopy and the fluorescence inverted microscope.The analysis of cell cycle and apoptosis of HepG2 was performed by flow cytometry.The RT-qPCR method was used to detect the changes in mRNA expression levels of genes involved in cell cycle and apoptosis.The results showed that the organic solvent extracts of Inonotus obliquus from Canada and Russia had higher inhibitory effects on both two human hepatocarcinoma cells,but they had highly toxic to the human normal liver cells.However,C-OWC,not only had a inhibition rate on both two human hepatocarcinoma cells,but also had no toxic effect on L02.And the inhibitory effect on HepG2 showed an obviously time-dose effect relationship,besides,the IC50 values were 1293.75 ?g/mL,581.64 ?g/mL,396.45 ?g/mL after 24,48,and 72 hours of administration at the concentrations from 62.5 ?g/mL to 4000 ?g/mL.The results of AO/EB staining showed that the cells appearred significant apoptosis characteristics which was showed lower green fluorescence and higher red fluorescence,increased membrane permeability,round cell,and increased apoptotic bodies after administration of C-OWC,The results of flow cytometry showed that the cell cycle was arrested in the G1 phase after administration of C-OWC,and the apoptosis rate was significantly increased compared with the control group.The results of RT-qPCR showed that the expression of Bcl-2 was decreased and the expression levels of Bax,Bid,and P21 were increased from the cells of the C-OWC group.In conclusion,C-OWC has certain anti-tumor activity by inhibiting the proliferation of HepG2 cells in human liver cancer,blocking cell cycle and inducing apoptosis.