Anti-hepatoma Effect Of Selaginella Granules And Exploration On Its Action Mechanism

Objective To study the inhibitory effect of selaginella granules to the growth of transplantation of hepatocellular carcinoma model,and to study its action mechanism by exploring the effect of selaginella granules to cell cycle-related protein,as well as tumor invision and migration-related protein.Method Hepatocellular carcinoma models were produced by injecting subcutaneously suspension of Hep G2 in Nu/Nu nude mice.A total of 48 successful Nu/Nu nude mice model of hepatocellular carcinoma were randomly divided into model control group,fluorouracil group,selaginella granules group,and combined treatment group.There were 12 mices in each group.Fluorouracil 20 mg/kg was given to mice in fluorouracil group by gastric gavage.Selaginella granules 20mg/kg was given to mice in selaginella granules group by gastric gavage.Fluorouracil 20 mg/kg and selaginella granules 120 mg/kg was given to mice in combined treatment group by gastric gavage.The same volume of pure water was given to mice in model control group.The duration in each group was 4 weeks.Tumor volume was determined in 2nd,3rd and 4th week,and the inhibitory rate of hepatocellular carcinoma volume.Mice were executed in 1st h after last treatment.Tumor tissue were separated and weighed carefully,and the inhibitory rate of hepatocellular carcinoma weight.RNA was extracted by Trizol from tumore tissue.m RNA of Cyclin D1,CKS1,FAK,Ezrin and MMP-9 was determined by RT – PRC.Protein of Cyclin D1,CKS1,FAK,Ezrin and MMP-9 was determined by western blotting.Results(1)Transplanted tumor volume in four groups was not significantly different before there was no statistical significance treatment(P > 0.05).Transplanted tumor volume in fluorouracil group was signifcantly decreasd compared to that in model control group in 1st,2nd,3rd and 4th week after treatment(P<0.01).Transplanted tumor volume in selaginella granules group was signifcantly decreasd compared to that in model control group in 2nd,3rd and 4th week after treatment(P<0.01).Transplanted tumor volume in combined treatment group was signifcantly decreasd compare to that in fluorouracil control group in 3rd and 4th week after treatment(P<0.01).(2)The inhibitory rate of hepatocellular carcinoma volume in combined treatment group was signifcantly decreasd compared to that in model control group in 1st,2nd,3rd and 4th week after treatment(P<0.01).The inhibitory rate of transplanted tumor volume in combined treatment group was signifcantly decreasd compare to that in selaginella granules group in 3rd and 4th week after treatment(P<0.01).(3)After 4 week treatment,m RNA expression of Cyclin D1 and CKS1 in fluorouracil group,selaginella granules group and combined treatment group was significantly decreased compared with that in model control group(P < 0.01),and m RNA expression of Cyclin D1 and CKS1 in combined treatment group was significantly decreased compared with that in fluorouracil group and in selaginella granules group(P < 0.01).(4)After 4 week treatment,protein expression of Cyclin D1 and CKS1 in fluorouracil group,selaginella granules group and combined treatment group was significantly decreased compared with that in model control group(P < 0.05 or P < 0.01),and protein expression of Cyclin D1 and CKS1 in combined treatment group was significantly decreased compared with that in fluorouracil group and in selaginella granules group(P < 0.05 or P < 0.01).(5)After 4 week treatment,m RNA expression of FAK,Ezrin and MMP-9 in fluorouracil group,selaginella granules group and combined treatment group was significantly decreased compared with that in model control group(P < 0.05),and m RNA expression of FAK,Ezrin and MMP-9 in combined treatment group was significantly decreased compared with that in fluorouracil group and in selaginella granules group(P < 0.05).(6)After 4 week treatment,protein expression of FAK,Ezrin and MMP-9 in fluorouracil group,selaginella granules group and combined treatment group was significantly decreased compared with that in model control group(P < 0.05),and protein expression of FAK,Ezrin and MMP-9 in combined treatment group was significantly decreased compared with that in fluorouracil group and in selaginella granules group(P < 0.05 or P < 0.01).Conclusion selaginella granules has anti-hepatoma effect,and it can enhance the curative effect of fluorouracil.Its anti-hepatoma effect may be connected with inhibition to cell cycle-related protein such as Cyclin D1 and CKS1,as well as to tumor invision and migration-related protein such as FAK,Ezrin and MMP-9.