论文部分内容阅读
AIM To screen and research targeting anti-hepatoma drugs from 18 kinds of the traditional Chinese herbal medicines from Yao medicin , Chuang medicine in Guangxi province by antitumor drug target as tyrosine kinase.METHODS 18 Ethanol extracts of Yao medicines , Chuang medicines in Guangxi were obtained by cold-extraction alcohol, then the inhibitory activity of human hepatoma cell line SMMC-7721 with 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-7721 at 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-7721 were established, and then observated the tumor growth by HE staining and expression of VEGF, CD34 in tumor tissues of each group by immunohistochemistry method.RESULTS Dysosma versipellis and Hudrocotyle sibthorpioides had stronger anti-tumor ability to human hepatoma cell line SMMC-7721 and BEL-7404 among the screening drugs, the IC50 of SMMC-7721 was 0.281 and 0.094 g· L-1 ; the IC50 of BEL-7404 was 0.235 and 0.104 g· L-1, respectively.Experimental results of serum pharmacology showed medicated serum inhibited the growth of human hepatoma cell line SMMC-7721 in varying degrees.10% serum of Dysosma versipellis group had an obvious inhibition effect on tumor compared with 10% serum of sorafenib group, the inhibitory rate was 11.15%.There was significant differences between two groups (P <0.05).Dysosma versipellis and Hudrocotyle sibthorpioides reduced the level of tyrosine kinase (P <0.01).Sorafenib 12.5 mg·L-1, Dysosma versipellis 50 g·L-1 and Hudrocotyle sibthorpioides 12.5-50 g·L-1 inhibited the angiogenesis of CAM in varying degrees.Compared with control group, serafenib had a significant inhibition on different levels of blood vessels (P <0.01).High dose of Dysosma versipellis had inhibition on two or three-level blood vessels (P < 0.01).Hudrocotyle sibthorpioides had 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 Dysosma versipellis group and medium-dose Dysosma versipellis group ,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 Dysosma versipellis group was significantly greater than control group.MVD were lower of sorafenib group and high-dosage Dysosma versipellis group than that of control group (P < 0.01).There were significant differences between medium-dose Dysosma versipellis group and control group (P < 0.05).Sorafenib group and high-dose Dysosma versipellis group reduced significantly the expression of VEGF in tumor tissues (P < 0.01).CONCLUSION Dysosma versipellis and Hudrocotyle sibthorpioides not 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 Dysosma versipellis.