| BackgroundLiver cancer is a serious threat to human health. In 2004, 610 000 people died of liver cancer. China is a high incidence region of liver cancer. At present, surgical treatment is still the main therapy, while the matters of palindromia and metastasis after surgery as well as most patients having missed the qualification for operation make pure surgical treatment cannot meet the present therapy on liver cancer. Traditional Chinese Medicine with its unique advantages has a long history on the treatment of tumor and has achieved a remarkable effect. Abstracting the effective components of traditional Chinese medicine and researching the pharmacological actions are important approaches for the studing of TCM treating tumor.Valvate actinidia root, a conventional medical material in the area of Jiangsu and Zhejiang provinces, is widely applied in the treatment of lung cancer and gastrointestinal tumors. There are about 20 kinds of mineral elements in valvate actinidia plant, rich of amino acid in the root, catechu, triterpenes and other components in ethanol extracts. Recent research proved that the AQ-type complex could inhibit the growth of mouse liver subcutaneous transplanted tumor. On the base of previous relative work, this subject have further observed the inhibitory action of total saponin from Actinidia Valvata Dunn on liver cancer in vivo and in vitro, and preliminarily discussed parts of its probably relevant mechanisms.ObjectivePART ONE:To observe the inhibitory action of total saponin from Actinidia Valvata Dunn on mouse liver subcutaneous transplanted tumor in vivo. PART TWO:To observe the impact of total saponin from Actinidia Valvata Dunn on the cell cycle of hepatoma carcinoma cell in vitro.MethodsPART ONE: experimental research on the inhibitory action of total saponin from Actinidia Valvata Dunn to the growth of liver cancer in vivo 1. Establishing mouse liver subcutaneous transplanted tumor model, random grouping, and regular administration. Weighing mice in each group before and after the experiment, measuring cubic capacity of the tumor, weighing the weight of the tumor;2. Immnohistochemically staining the tumor issues, applying IMS medical figure quantitative analysis software to detect the expression of tumor issue TUNEL.PART TWO: experimental research on the effect of total saponin from Actinidia Valvata Dunn to liver cancer in vitro.1. Detecting the growth inhibitory effects of different concentrations of total saponin from Actinidia Valvata Dunn to different periods of liver cancer cell strain BEL-7402, H22;2. Detecting the cycle distribution impact of different concentrations of total saponin from Actinidia Valvata Dunn effecting 48 h on liver cancer cell strain BEL-7402, H22 with PI staining method by FCM3. Detecting the changes of the cell cycle relevant protein and kinase of human liver cancer cell strain BEL-7402 after being acted by total saponin from Actinidia Valvata Dunn with Western blot method.ResultPART ONE: experimental research on the inhibitory action of total saponin from Actinidia Valvata Dunn to the growth of liver cancer in vivo1. No obvious difference on the weight of each groups of mice; the tumor size and weight of the group on drug are obviously different from those of the blank group, there's significant statistic difference (P<0.05); The tumor inhibitory rate of the high dose group and the lower dose group are respectively 53.2% and 43.5%.2. The masculine expression rate of TUNEL is higher than the blank group, and has statistic significance (P<0.05).PART TWO: experimental research on the effect of total saponin from Actinidia Valvata Dunn on liver cancer in vitro1. The result of MTT and Trypan blue shows that there is dose dependence for the growth inhibition action of total saponin from Actinidia Valvata Dunn on human liver cancer cell BEL-7402. There's no obvious cytotoxic action when the concentration of total saponin from Actinidia Valvata Dunn is lower than 0.75 mg/ml; no obvious cytotoxic action was seen when applying total saponin from Actinidia Valvata Dunn on the concentration of 0.75 mg/ml to mouse liver cancer cell H22.2. When total saponin from Actinidia Valvata Dunn acting on human liver cancer cell BEL-7402, the proportion of S period significantly increased with the increase of administration concentration (0.1 mg/ml, 0.2 mg/ml, 0.4 mg/ml), while the proportion of the other two periods decreased. When total saponin from Actinidia Valvata Dunn acting on mouse liver cancer cell H22, for H22 the proportions of G0/G1 period and G2/M period significantly increased with the increase of administration concentration (0.2 mg/ml, 0.4 mg/ml), while the proportion of S period decreased.3. According to the increase of the administration concentration of total saponin from Actinidia Valvata Dunn the expression of cell cycle relevant protein Cyclin A of BEL-7402 was up-regulated.Conclusion1. Total saponin from Actinidia Valvata Dunn could inhibit the growth of mouse liver subcutaneous transplanted tumor, enhance the expression of TUNEL, one of its related action mechanisms on inhibiting mouse subcutaneous transplanted tumor might be necrosis promotion to the tumor.2. At high concentration, total saponin from Actinidia Valvata Dunn inhibits the proliferation of human liver cancer cell BEL-7402 through cytotoxic action; at low concentration, it blocks the cell cycle of human liver cancer cell BEL-7402 in S period, its action mechanism of cell cycle blockage might be related with the up-regulation of the expression of cell cycle relevant protein Cyclin A.3. On obvious cytotoxic action was seen when total saponin from Actinidia Valvata Dunn n acting on mouse liver cancer cell H22 at the concentration of 0.4 mg/ml.4. The cell cycle of mouse liver cancer cell H22 could be blocked in periods of G0/G1 and G2/M by the action of total saponin from Actinidia Valvata Dunn... |
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