The Anticancer Molecule Mechanism And The Basic Experiment Study In Prevention Of Hepatocellular Carcinoma With Green Tea Tolyphenols

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the world. The research on prevention and treatment of HCC is the most challenging topic .Green tea and its key component green tea polyphenols (GTP) have been paid great attention for its anticancer effects.GTP has extensive biologic features and pharmacology effects. It can inhibit the genesis and development of many tumor. But the exact mechanism of the GTP remains unknown. This study explore the anticancer effects of GTP through three following experiments ,they are cell experiments in the vitro, nude mice mode of HCC cell line SMMC-7721 and the basic experiment study in prevention of HCC with GTP. DNA agarose gel electrophoresis, Flow cytometer(FCM), the examination of telomerase, Atlas cDNA array, RT-PCR and electron microscope, immunohistochemistry were used in the study.In vitro cell experiments ,MTT and colony formation experiments show that GTP can apparently inhibit the proliferation and colony formation of HCC cell line Hep3B,with the increase in GTP's concentration, its anticancer effects augments .the GTP's IC50 is 143μg/ml. After being processed with GTP, the shape of HCC cell changes. Through electron microscope ,apoptosis of cell can be watched. The cell DNA extracted display ladder bands in DNA agarose gelelectrophoresis. Through the analysis of flow cytometry.,GTP can block cell cycle Gi, preventing the damaged cell entering S phase ,so it can suppress the excess proliferation ,GTP can induce the apoptosis of Hep3B. It showed dose -time and time -effect relations . When Hep3B cell was induced by GTP ,The telomerase activation decrease . Atlas cDNA array was conducted .The results showed that GTP can upgrade related gene TNF,TNFR,Caspase -9 and Caspase-10 which can induce apoptosis while down regulating bcl-2. RT-PCR confirm the result which is in accordance with the result of Atlas cDNA array.Establishing the transplantation tumor in nude mice using HCC cell line SMMC-7721. The studies showed that GTP can inhibit the transplantation tumor in nude mice. Through electron microscope ,we can see that GTP can induce the cell apoptosis of the transplantation tumor in nude mice .Through flow cytometry ,the apoptosis rate in positive group and high ,medium, low dose group of GTP is much higher than that from control group.and the difference is significant (P<0.05). In the experiment of immunohistochemistry,the protein expression of gene Bcl-2 in positive group and high ,medium ,k>w dose group of GTP decrease compared with that from control group . Also the difference is significant(P<0.05)., No difference was found in the protein expression of gene 53 in transplantation tumor of negative control group and high ,medium ,low dose group of GTP (P>0.05).The basic experiment study in prevention of HCC with GTP assess the effects of Long-tern taking GTP on the immune functions and anti-oxidation among people with high risks of suffering from HCC. 1200 people with high risks of...