Screening And Research Selective Anti-Hepatoma Drugs From The Traditional Chuang And Yao Medicine In Guangxi By Tyrosine Kinase, Angiogenesis As Antitumor Drug Targets

Objective:To screen and research targeting anti-hepatoma drugs from18kinds of the traditional Chinese herbal medicines from Yao medicin,Chuang medicine in Guangxi by antitumor drug target as tyrosine kinase.Methods:18Ethanol extracts of Yao medicines,Chuang medicines in Guangxi were obtained by cold-extraction alcohol,then the inhibitory activity of human hepatoma cell line SMMC-7721with MTT were determined in vitro.The medicated serums with obviously anti-tumor effect of enthanol extracts were prepared according to the serum pharmacology method and further observed its inhibiting effect in same tumor cell line by MTT.The total protein of human hepatoma cell line SMMC-7721at different time or concentration of extracts was extracted by cell lysis buffer,respectively.The content of tyrosine kinase in total protein of tumor cells was detected by Enzyme-linked immunosorbent assay (ELISA). The model of angiogenesis in allantocheriionof chick embroyo was established so as to observe antiangiogenesis effect of extracts by CAM assay in vivo.The models of xenografted tumor in nude mice with human hepatoma cell line SMMC-7721were established,and then observated the tumor growth by HE staining and expression of VEGF,CD34in tumor tissues of each group by immunohistochemistry method.Results:Yao medicine2and Yao medicine5had stronger anti-tumor ability to human hepatoma cell line SMMC-7721among the screening drugs,the IC50was0.080mg/ml,0.281mg/ml,respectively.Experimental results of serum pharmacology showed medicated serum inhibited the growth of human hepatoma cell line SMMC-7721in varying degrees.10%serum of Yao medicine2group had an obvious inhibition effect on tumor compared with10%serum of sorafenib group, the inhibitory rate was11.15%.There was significant differences between two groups (P<0.05).Yao medicine2and Yao medicine5reduced the level of tyrosine kinase (p<0.01).12.5mg/L sorafenib,50mg/mL Yao medicine2and12.5mg/mL-50mg/mL Yao medicine5inhibited the angiogenesis of CAM in varying degrees.Compared with control group,sorafenib had a significant inhibition on different levels of blood vessels(p<0.01).High dose of Yao medicine2had inhibition on two or three-level blood vessels (p<0.01).Yao medicine5had inhibition on three-level blood vessels,while inhibiting one or two-level blood vessels under high dosage (p<0.01).The tumor weight and tumor relative volume in the following groups,such as sorafenib group,high-dose Yao medicine2group and medium-dose Yao medicine2group,was significantly lower than control group(P<0.01). The result of HE staining showed that tumor necrotic area in sorafenib group and high-dosage Yao medicine2group was significantly greater than control group.MVD were lower of sorafenib group and high-dosage Yao medicine2group than that of control group (P<0.01).There were significant differences between medium-dose Yao medicine2group and control group (P<0.05). Sorafenib group and high-dose Yao medicine2group reduced significantly the expression of VEGF in tumor tissues (P<0.01).Conclusion:Yao medicine2and Yao medicine5not only can inhibit the growth of blood vessels in varying degrees,but also can reduces the content of tyrosine kinases in tumor cells.In addition,MVD and the expression of VEGF may be reduced because of Yao medicine2.