Study On Inhibitory Effects And Machanism Of Ezhukuijian Decotion On Human Pancreatic Cancer

Objective To study on inhibitory effects and machanism of Ezhukuijian Decotion on human pancreatic cancer and explore the potential mechanism of Ezhukuijian Decotion in the treat ment of pancreatic cancer.Methods:The expression of DcR3protein in pancreatic carcinoma tissue chips were detected by IHC; The expression of DcR3mRNA were detected by real-time PCR in pancreatic cancer cell lines AsPC-1, BxPC-3, PANC-1, MiaPaCa-2. BxPC-3and MiaPaCa-2cell lines were chosen for the following study.Three different small interfering RNA for human DcR3(siRNAl, siRNA2and siRNA3) and one negative siRNA were designed.The expression of DcR3after DcR3RNAi or negative RNAi were identified by real-time PCR. Cell growth,migration, apoptosis and invation were detected respective by MTT, wound healing assay,FCM and in vitro invasion assay. Then investigate the mechanism of inhibitory effects of Ezhukuijian Decotion on pancreatic cancer cell lines.To examine the effects of Ezhukuijian Decotion on tumor growth,we performed the in vivo assay using an subcutaneous transplanted tumor model in the nude mice.Then human pancreatic cancer line MiaPaCa-2burdened nude mice were randomized to receive Ezhukuijian Decotion at different doses and oral chemotherapeutics, Inhibitory rates of tumor growth in vivo were calculated,and the expression of DcR3protein was detected by western blot.Results:The expression of DcR3protein was higher in pancreatic cancer tissues than in normal pancreatic tissues. The expression of DcR3was positively realated Pathological staging.The expression of DcR3were dected in the four human pancreatic cancer cell lines, the level of DcR3expression was higher in MiaPaCa-2and BxPC-3. BxPC-3and MiaPaCa-2cell lines were chosen for the following study,The expression of DcR3mRNA in DcR3RNAi transfectants was21%,24%and28.3%respectively of that in control RNAi transfectants. The DcR3-siRNA3and negative-siRNA were selected for the subsequent studies. We found that Ezhukuijian Decotion inhibited pancreatic cancer cell lines cell growth,migration,apoptosis and invation abilities. Ezhukuijian Decotion could inhibit the growth of human pancreatic cancer cell line MiaPaCa-2in vivo.Tumor weight in Ezhukuijian Decotion groups was sithificantly lower than that in conrol group.The expression of DcR3protein in Ezhukuijian Decotion group was significantly lower that in the control group.Conclusions:Ezhukuijian Decotion may remarkablely inhibit the growth of human pancreatic cancer cell in vivo and vitro.The potential mechanism is to decrease DcR3expression through inhibiting MAPK sigaling pathway.