Establishment Of A Human Hepatocellular Carcinoma Model In Nude Mice And Evaluate Matrine To Enhance The Cisplatin’s Anticancer Effect For Transplantation Tumor And Its Influence On The Expression Of Survivin/Caspase-3

Objective: To establish a human hepatocellular carcinoma model in BALB/c nude mice and to investigate the effect on the expression of survivin and caspase-3 of cisplatin(cisplatin, DDP) combined with Matrine(Matrine, Ma) on nude mice xenograft model of human hepatocellular carcinoma, and clarify the potential molecular mechanism. Methods: Twenty four 6-week old male BALB/c nude mice were used in this study. A xenograft model of human hepatocellular carcinoma was established by subcutaneous injection of Hep G2 cells into the axilla in nude mice. Then the nude mice were randomly divided into 6 groups: control group, Ma group(a 3-wk group, a 2-wk group, a 1-wk group), DDP group and DDP combined with Ma group and the all nude mice were injected with experimental drug via intra peritoneal injection. The general status and body weight of mice, the time to tumor formation, tumor volume and weight, the nude mice’s weight change curve and the tumor growth curve were assessed. After 14 days drug intervention, the experimented nude mice were sacrificed and the tumor tissue were got to be weighed and the tumor inhibition rate was counted, and pathological changes were observed after HE staining toevaluate the anticancer effect of matrine. The expression of survivin and caspase-3 were detected by immunohistochemistry method. Results: The success rate of establishing the xenograft model was 100%. Matrine had an inhibitory effect on the growth of tumor xenografts. The reduced rates of tumor growth was 8.3% in the 1-wk group, 37.5% in the 2-wk group, and 54% in the 3-wk group.The reduced rates of tumor growth were significantly higher in the 3-wk and 2-wk groups than in the 1-wk group and control group(P < 0.01). There was also a significant difference between the 1-wk group and control group(P < 0.05), and between the 3-wk group and 2-wk group(P < 0.05). The tumor inhibition rate is respectively : Ma group 37.5%, DDP group 75%, DDP combined with Ma group 83.3%. DDP combined with Ma can significantly inhibit the tumor growth compared with Ma group and DDP group(P < 0.05), It was also found that the average weight of nude mice in Ma+DDP group is 21.5g, lower than NS group(28.5g) and Ma group(26.67g), however, more than DDP group(17.33g). what’s more, the whole body situation, containing spirit, action, appetite etc, is also better than DDP group. No obvious differences were found in histology of cancerous tissues between the five treatment groups and the control group, the only different was that the treament groups showed some apoptotic cells and coagulation necrosis. In tumor tissue, the positive expression rate of Survivin in Ma+DDP group is:(19.58±4.52)%, lower than NS group:(83.26±15.56)%, Ma group: 1-wk group(79.58±16.36)%, 2-wk group( 62.50±8.09) %, 3-wk group( 55.56±8.16) %, and DDP group:(38.67±8.26)%(P<0.05). To the contrast, The positive expression rate of caspase-3 in NS group, Ma 1-wk group,2-wk group,3-wk group and DDP group are respectively:(21.15±3.68)%、(24.65±2.29)%、(35.13±10.57)%、(43.68±10.96)%、(65.88±4.81)%, but that in Ma+DDP group(78.26±6.09)%is clearly higher than the others. Ma+DDP group showed significantly different results for the others(P<0.05). Conclusions: Matrine has anticancer effect on nude mouse xenograft model of human hepatocellular carcinoma and without obvious toxic effects, and early drug intervention has more obvious effect than later. Combined with DDP, Ma can enhance DDP’s anticancer effect for hepatocellular carcinoma nude mice xenograft, improve the whole body status, reduce DDP’s toxicity, and its effect mechanism may be influencing the expression of Survivin/caspase-3 and inducing the tumor cells apoptosis.