| Objective:By adopting the method of cell culture in vitro and animal experiments, observing the effect of Eucheuma seaweed pigment glycoprotein (SPG) on the proliferation of mice hepatocarcinoma cells, the expression level of PCNA and Bcl-2 protein of tumor tissue, the inhibition rate and other indicators.Methods:1 In vitro experiment:Different concentrations of SPG (100,50,10mg /L) were cultured with logarithmic phase of H22 hepatocarcinoma cells of mice for 24 hours. Distilled water was set as a separate control group, 20mg/L cyclophosphamide as a positive control. The cell proliferation was determined by MTT assay.2 In vivo experiment:A total of 50 healthy Kunming mice of half gender, weighing 18-20 g, adaptively fed in the laboratory for 5 days. Under sterile conditions, the H22 passage section 6 days of the mouse ascites liver cancer was inoculation to the healthy mice, which were inoculated subcutaneously at right axilla 0.2 ml. All of the animals were randomly divided into five groups after inoculation 24 hours, which were tumor control group, high, medium and low doses of seaweed pigment glycoprotein group and cyclophosphamide group with 5 males and 5 female mice in each group. Mice in high, medium and low dose group were given by gavages concentration of 100,50 and 10 mg/kg seaweed pigment glycoprotein aqueous, tumor control group were treated with saline, once a day for 10 days, and cyclophosphamide 20mg/kg was injected to the cylophosphamide group every the other day. Mice in each group ate and drank freely. The total of the mice were killed after the last administration. The proliferation activity of hepatocarcinoma cells was determined by MTT assay, the level of PCNA and Bcl-2 protein was measured by immunohistochemistry method, and tumor weight was examined by electronic scale and calculated the inhibitory rate.Results:1 In vitro experiment results:The proliferative activity of cultured cells in vitro in 100 mg/L seaweed pigment glycoprotein group were (0.404±0.008), which were significantly lower than that in tumor control group(0.545±0.002) (P＜0.05), and those in 50 mg/L and 100 mg/L seaweed pigment glycoprotein group were (0.445±0.005) and (0.515±0.009) respectively. Indicates that a certain dose of seaweed pigment glycoprotein has inhibition effect on proliferation activity of H22 hepatoma cell, and with increasing doses of SPG, proliferation activity of H22 hepatoma cell was decreased (P<0.05).2 In vivo experiment results:(1) The proliferative activity:The cell proliferation activity expressed by the optical density of the high-dose group and tumor control group were (0.711±0.028) and (1.135±0.032), respectively (P＜0.05).(2) The level of PCNA protein:The positive expression rate in high dose group were 28.32%, and those in tumor control group were 72.78%, the differences were significant (P＜0.05).(3) The level of Bcl-2 protein:The positive expression rate in tumor control group and high dose group were 65.16% and 16.78% respectively, the differences were significant (P＜0.05).(4) The inhibition rate of tumor growth:The average of tumor weight in tumor control group and high dose group were (0.55±0.23) g and (0.31±0.05) g respectively, the differences were statistically significant (P＜0.05). The tumor growth inhibition rate in high dose group was 43.64%.Conclusion:1 Seaweed pigment glycoprotein (SPG) has inhibition effect on the proliferative activities of H22 hepatoma cells.2 Seaweed pigment glycoprotein (SPG) could reduce the expression level of PCNA protein on H22 hepatoma tissue, inhibit proliferation of hepatoma cells.3 Seaweed pigment glycoprotein (SPG) could reduce the proliferative activities of H22 hepatoma cells, reduce the expression of Bcl-2 protein on H22 hepatoma tissue.4 Seaweed pigment glycoprotein (SPG) can inhibit growth of hepatoma tumor on mice, which provide scientific basis on the prevention and treatment of liver diseases. |
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