The Effects Of β-catenin In BMSC Participate Hp Chronic Infection Induced Gastric Cancers

Aims:Wnt signaling is involved in the process of gastric cancer,the P-catenin/Tcf4/Fzd5is a key protein in the classic Wnt signaling pathway,BMSC are thought to be involved in gastric cancer,but the mechanisms are unclear.This topic takes Wnt signaling pathway as a target, by the technology of shRNA interference the mouse bone marrow stromal stem cells(BMSC) and determinating the P-catenin protein expression of BMSC,to explore the mechanism of BMSC in Hp chronic infection induced gastric cancers after β-catenin-shRNAtransfected BMSC.Methods:Building the specific eukaryotic expression vector containing the β-catenin-shRNA. BMSC with a10%FBS-containing DMEM culture medium, according to a concentration of2×105/well seeded in a Transwell upper chamber, incubated for24h in37℃and50mL/L CO2incubator.When the cell adherent rate of60%,transfected with the Lipo-fectamine2000.Take the P-catenin-shRNA treated as experimental group, shRNA missense sequence treated as a negative control group. Determine the efficiency of the transfected BMSC,After48h,detected the P-catenin expression of BMSC by Western Blot. Proportion of BMSC, mice gastric cancer cell (MFC), HP sonication matter (1:10:100) to establish Transwell in vitro co-culture system. Incubate the co-culture system for24h in the incubator, Detected the β-catenin mRNA and protein changes in the MFC by Real-time PCR method and Western Blot. Results:β-catenin-shRNA transfected BMSC after48h, the β-catenin protein in the BMSC was decreased. BMSC、MFC、Hp co-cultured for24h, Compared with the negative control group, the β-catenin mRNA in MFC expression was significantly decreased (0.488±0.120;1.026±0.057; P<0.05), the P-catenin protein expression was also decreased.(0.687±0.107;1.06±0.113; P<0.05).Conclusion:BMSC are thought to be involved in the process of Hp chronic infection induced gastric cancers early event in the Wnt signaling pathway. BMSC as a genetically engineered cells, to provide new ideas and methods for the treatment of gastric cancer in the future.