Research On Tumor Prevention Effects Of ATCan In Mouse Primary Liver Cancer Models Induced By DEN Treatment

Hepatocellular carcinoma(HCC)is one of the most frequent malignancies worldwide.lt is the fifth most common kind of tumours and the third most common cause of death among cancer patients..The HCC incidence rate is one million cases every year,More than 600,000 people die from HCC each year..In addition,the vast majority of patients present to the clinic with advanced stage disease,these patients typically die within three to six months as a result of lack of effective treatment and their five-year survival is only 14%.Worldwide research on the disease needs to be intensified in both the medical and pharmaceutical fields.N-diethylnitrosamine(DEN)is one of the most important environmental carcinogens,which is widely present in many foods,such as alcoholic beverages,cheese,processed meat,soybeans,tobacco products,cosmetics and agrochemicals.These products will increase the risk of liver cancer in humans.Therefore,DEN-induced liver cancer model mice are widely used in liver cancer studies.Altered metabolism is a universal property of most cancer cells.Compared to normal cells,cancer cells strongly upregulate glucose uptake and glycolysis to give rise to increased yield of intermediate glycolytic metabolites and the end product pyruvate.Finally,following glycolysis,most pyruvate is converted to lactate in the cytoplasm by the action of lactate dehydrogenase(LDH)and secreted,rather than being oxidized through mitochondrial metabolism.The metabolic phenomenon is referred to the "Warburg effect".The alteration of glycolysis has been recognized in recent years as an emerging hallmark of cancer.ATCan is a processed ginseng product,which is produced by our laboratory.In this study,we focus on its role in the prevention of hepatocellular carcinoma,and try to clarify its mechanism.For that,the following researches been done:1)We performed the High performance liquid chromatography(HPLC)to detect the content of ginsenosides in ATCan and wild ginseng.We found that ATCan is rich of ginseng saponins,and ginsenoside Rkl,Rg3,Rg5 are the maximurm contents;2)We have established a DEN-induced C57/BL6 mouse model of hepatocellular carcinoma,by intraperitoneal injection of DEN(25mg/kg)in mice at the 15th day after birth.After that,mice were randomly divided into four groups,namely,model group,low-dose ATCan group(50mg/kg),high-dose ATCan group(100g/kg)and wild ginseng group(1OOmg/kg).Mice were administered orally at 8 weeks of age,once every two days,which is last for 10 weeks;3)When the mice were sacrificed at 8 months of age,the incidence of cancer and tumor size are detected in each group of mice.We observed the livers and tumors through the naked eye,performed HE staining of the livers to observe the pathological changes in mice.We found that ATCan and wild ginseng significantly reduced tumor incidence of DEN-induced C57/BL6 mice:the occurrence of tumors in mice model group was 44.44%(4/9);the incidence of hcc in the low-dose ATCan group,the high-dose ATCan group,and the wild ginseng group were 7.69%(1/13),7.14%(1/14)and 6.67%(1/15)respectively.This result shows that both ATCan and wild ginseng play important roles in the prevention of DEN-induced C57/BL6 hcc;4)To clarify the mechanism of the prevention of liver cancer of ATCan and wild ginseng,we examined the effect of different concentrations of wild ginseng and ATCan to human hepatoma cells SK-HEP-1 on glycolysis.We found that ATCan has a significant dose-dependent inhibitory effect on glycolysis of SK-HEP-1.This result suggests that the antihepatoma effects of ATCan is probably by inhibiting glycolysis;5)We examined the glycolysis with the treatment of the major ginsenosides of ATCan on SK-HEP-1 cells.The results show that ginsenoside Rkl,Rg3,Rg5,Rk3,Rh4 can inhibit the glycolysis of SK-HEP-1 cells.Therefore,we speculate that the ginsenoside components in ATCan may play important roles in the prevention of hepatocellular carcinoma.Taken together,ATCan can prevent DEN-induced C57/BL6 hepatocellular carcinoma,and at least part of this effect is achieved by the inhibition of glycolysis which is owe to the ginsenoside components of ATCan.