This topic takes Wnt signaling pathway as a target, by the technology
of shRNA interference the mouse bone marrow stromal stem cells (MSCs) and determinating the β-catenin protein expression of MSCs, to explore the mechanism of MSCs in Hp chronic infection induced gastric cancers after β-catenin-shRNA transfected MSCs. Methods: Building the specific eukaryotic expression vector containing the β-catenin-shRNA. MSCs with a 10% FBS-containing DMEM culture medium, according to a concentration of 2 × 105/well seeded in a Transwell upper chamber, incubated for 24 h in 37°C and 50 mL/L CO2 incubator. When the cell adherent rate of 60%, this website transfected with the Lipo-fectamine 2000. Take the β-catenin-shRNA treated as experimental group, shRNA missense sequence treated as a negative control group. Determine the efficiency of the transfected MSCs, After 48 h, detected the β-catenin expression of MSCs by Western Blot. Proportion of MSCs, mice gastric cancer cell (MFC), HP sonication matter (1 : 10 : 100) to establish Transwell in vitro co-culture system. Incubate the co-culture system for 24 h 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 MSCs after 48 h, the β-catenin protein in the MSCs was decreased about 25%. MSCs, MFC, Hp co-cultured for 24 h, Compared with the negative control group, the β-catenin mRNA in MFC expression was significantly decreased (0.65 ± 0.30; selleck screening library P < 0.05), the β-catenin protein expression was also decreased (1.09 ± 0.06; P < 0.05). Conclusion: MSCs are thought to be involved in the process of Hp chronic infection induced MCE公司 gastric cancers early event in the Wnt signaling pathway.
MSCs as a genetically engineered cells, to provide new ideas and methods for the treatment of gastric cancer in the future. Key Word(s): 1. β-catenin; 2. Wnt signal; 3. RNA interference; 4. MSCs; Presenting Author: JING LV Additional Authors: XU SHU, JIAN YI, NONGHUA LV Corresponding Author: XU SHU Affiliations: Nanchang University First Affiliated Objective: To investigate the relation between cell injury and repair in different Hp concentration and duration of infection in gastric mucosal, further provide a theoretical basis for its Hp pathogenic mechanisms. Methods: Selecting Hp ACTC43504 (CagA+, VacA+) standard strains infect GES-1 cell, Use logarithmic growth phase of Hp infection GES-1, take bacterial cells ratio 100 : 1, Hp infect gastric GES-1 0 h, 1 h, 3 h, 6 h, 12 h, 24 h; the different Hp concentration infect GES-1 cell in 6 h, bacterial cell ratio choose 0.50 : 1, 100 : 1, 150 : 1, 200 : 1, 300 : 1, then detect: DNA damage, ROS level, APE-1 expression, APE-1 intracellular localization.