| Objective：This study used Lentivirus to disturb the β-catenin signaling pathway in humanAGS and MGC-803gastric cancer cells, then detected the differential expression ofmicroRNAs (miRNAs) before and after interference via miRNA microarray. Thestudy makes it possible to search the mechanisms of β-catenin signaling pathwayand related miRNAin regulating gastric cancer.Methods：To obtain the stable transfected gastric cancer cells, lentivirus vectors targetingβ-catenin gene (ctnnb1)were designed to infect AGS and MGC-803cell linesrespectively. Then using real-time PCR and Western Blot to test the inhibition ratio。AGS cells were divided into two groups: AGS interference group and AGS controlgroup. MGC-803cells were also divided into two groups: MGC-803interferencegroup and MGC-803control group. Using miRNA microarray to screen thedifferential expression of microRNAs before and after β-catenin inhibition in eachcell line. Part of the miRNAmicroarray results were validated by real-time PCR.Results：1. Two lentivirus, LV3-CTNNB1-homo-433and LV3-CTNNB1-homo-769,were obtained. LV3-CTNNB1-homo-433could suppress β-catenin expression bothin AGS cells and MGC-803cells, LV3-CTNNB1-homo-769could suppress β-catenin expression only in MGC-803cells, and had no effect inAGS cells.2. miRNA expression profiles changed obviously afterLV3-CTNNB1-homo-433infection both in AGS cells and MGC-803cells. Comparedwith the control group, there were40miRNAs up-regulated and32miRNAsdown-regulated in AGS interference group, there were56miRNAs up-regulated and86miRNAs down-regulated in MGC-803interference group. The result ofhsa-miR-135b-5p and hsa-miR-210validated by Real-time PCR were consistent withthe results of the miRNAmicroarray.Conclusion： 1. Interference of β-catenin expression in human gastric cancer cells AGS andMGC-803influence the expression of microRNAs.2. miR-135b-5pã€miR-210are like to be regulated by β-catenin signalingpathway to play roles in gastric cancer. |
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