Inhibition Of Mung, Gin And L08 To The Proliferation Of Human Liver Cancer Cell SMMC7721 And The Expression Of CD133 In Cancer Stem Cell

Background and objective:Liver cancer is one of the most popular malignant tumor. Its mortality in malignant tumors of the digestive system was third.In China, about 110 thousands of persons died of liver cancer each year. Main reason to lead to high mortality of liver cancer is the relapse and transfer. Why can malignant tumor recure and transfer, and be resistant to the therapies of radiation and chemicals? It has been discovered in new study that there are some cells to be resistance to drugs in tumor tissue, which are known as cancer stem cells(CSCs) or tumor stem cells(TSCs). These cells are very little in tumor tissue. About amount is 1% in total tumor cells. However, they can differentate into tumor cells and continuously proliferate. It has been discovered that there exist some markers in tumor stem cells, and CD133 protein is a marker which extensively locates in the surface of tumor stem eels. Many studies have proved that there is a close relationship between CD133 protein and the occurance, development, invasiveness and transfer and produce of drugresistance of tumor cells. Cancer stem cells not only exist in tumor tissue, but also in cell lines which can becultured in vitro. Up till now, the function of CD133 protein is not too clear, but it has an important value in the fields to trace the existence of tumor stem cells in tumor tissue and to study the relationship between the expression level of CD133 protein and the occurance, development, invasiveness and transfer and produce of drugresistance of tumor cells. The hexokinase is an enzyme to phosphorylate six-carbon sugar, especial glucose. In most tissues and organisms, the phosphorylation of glucose is first step of the activation of glucose metabolism. Alkaline phosphatase is a hydrolase to be responsible for removing phosphate groups from many types of molecules, including nucleotides, proteins, and so on. In tumor tissues, the activities of hexokinase and alkaline phosphatase go up.Mung, Gin and L08 are some compounds of flovones and triterpens from mungbean, ginger and ganoderma. In this experiment, the inhibitions of Mung, Gin and L08 to the proliferation of human liver cancer cell SMMC772 and to the expression level of CD133 protein in liver tumor stem cells have been preliminarily studied, so that can supply experimental basis for the prevention of the relapse of liver cancer.Materials and methods:1. Culture liver cancer cell SMMC7721 to go in logarithmic growth time.2. Prepare 100μg/ml,200μ,/ml,300μg/ml,400μg/ml,500μg/ml of solutions of Mung, Gin and L08, and treat liver cancer cell SMMC7721 in logarithmic growth time for 24h,48h and 72h, and detect the activity of the cells by MTT method, and draw growth curve the cells.3. Treat liver cancer cell SMMC7721 with Mung, Gin and L08, and measure the activities of hexokinase and alkaline phosphatase in liver cancer cell SMMC7721 by the kits of hexokinase and Alkaline phosphatase.4. Treat liver cancer cell SMMC7721 with Mung, Gin and L08, and detect the expression level of CD133 protein in human liver cancer stem cells by Western blot method.Result:1. At experimental concentration, Mung, Gin, and L08 have inhibitions to human liver cell SMMC7721 after the treatment of the drugs, and there is close relationship between drug concentration and inhitions.2. At experimental concentration, Mung, Gin, and L08 have inhibitions to the activities of hexokinase and alkaline phosphatase in liver cancer cell SMMC7721.3. At experimental concentration, Mung and L08 have clear inhibitions to the expression level of CD133 protein, and there is a close relationship between drug dosages and the expression levels of CD133 protein. But, Gin does not lower the expression levels of CD133 protein.Conclusion:1. Mung, Gin and L08 have the inhibitions to the proliferation of human liver cancer cell SMMC7721, and this inhibition is related to the dosages of these drugs.2. Mung. Gin and L08 can inhibit the activities of hexokinase and alkaline phosphatase in human liver cancer cell SMMC7721.3. Mung and L08 can inhibit the expression of CD133 protein in human liver cancer stem cells. But, Gin can't lower the expression level of CD133 protein.