Study On Puerarin Injection Reverse Multidrug Resistance Of Human Gastric Cancer

Objective To find out a new MDR reversal medicine.We observe the reversal effect of puerarin on human muitidrug-resistant gastric carcinoma cell line SGC7901/VCR.We transplanted orthotopically tissue of human gastric carcinoma SGC7901/VCR into nude mice stomach wall to observe the reversion effect of puerarin on multidrug resistance of nude mice model of human gastric carcinoma in vivo and preliminarily investigate its reversal mechanism.Methods The cell cycle alteration,intracellular drug concentration of these cells,the sensitivity of gastric cancer cells to several anti-tumor drugs and expression of P-gp and MRP in gastric cancer cells were detected by FCM,MTT assay and immunocytochemical staining,respectively. Reinforcement microscope show Fluorescence intensity.The study was carried out on the 50 BALB/c nude mice models,transplanted orthotopically tissue of human gastric carcinoma SGC7901/VCR into their stomach wall by biologic mucus gluing technique.After 24h of establishment models,they were divided randomly into five groups:normal saline group,5-Fu group, 5-Fu+puerarin group,5-Fu+verapamil group and puerarin group,by celiac injection for thsree weeks.The inhibitary rates of tumor growth were detected,and local invasiveness,the rates of metastasis in the local lymph node,liver and pleen were compared among different groups after 10 weeks.The expressions of P-gp and MRP were detected by immunohistochemical technique.Results Puerarin injection could reserve the multidrug resistance(MDR) of SGC7901/VCR to 5-Fu,VCR,DDP and ADM.By comparing with SGC7901/VCR , SGC7901/VCR with P-gp and MRP cells decrease in the G1 stage and increase in the S and G2+M stage,and Fluorescence intensity was reinforced obviously. P-gp and MRP expression level was lower than that without puerarin injection.And then we successfully transplanted orthotopically tissue of human gastric carcinoma SGC7901/VCR and SGC7901 into nude mice stomach wall and establish an orthotopic model of human MDR gastric carcinoma.The transplanted carcinoma are similar with human gastric carcinoma between configuration and biology.There are stillMDR characteristic of tissue cell to 5-Fu,VCR,DDP and ADM,examining by MTT assay.There is no significant difference compared witn that before transplanting.The expression of P-gp was found in two guoups.But the expression of P-gp was +-++ in SGC7901 group,and +++ in SGC7901/VCR group.Detected by pathology tissue technique,all group were found lymph nodes metastasis.The rates of visceras metastasis in chemotherapy group was lower significantly than that in normal saline group(P<0.05),but no difference was found between puerarin group and normal saline group.The inhibitary rates of puerarin+5-Fu group and 5-Fu group were 56.69% and 18.11%,compared each other,P<0.05.P-gp and MRP expression level of puerarin+5-Fu group was lower than of the 5-Fu group.The rates of metastasis in the local lymph node,liver and pleen were not significant difference between them.The direct cytotoxicity to tumors,the rates of visceras metastasis were not significant difference between puerarin group and normal saline group.But P-gp and MRP expression level of puerarin group was significantly lower than of the normal saline group(p<0.05).Conclusion Puerarin injection could reserve the multidrug resistance(MDR) of SGC790I/VCR and nude mice model of human gastric carcinoma constructed using orthotopic,tbr which the decreased expression of P-gp and MRP maybe responsible.