Interference Of ShRNA On β-catenin Of CD138~- Cells In U266 Multiple Myeloma Cell Lines

Objective To explore the expression of β-catenin in Wnt1/ β-catenin signaling pathway of U266 multiple myeloma cell line and CD 138" cells in the line.Investgate the interference in expression of β-catenin with shRNA special for β-catenin-in CD 138- by observe CD 138" cell growth changes and expression levels of β-catenin mRNA and P-catenin protein of it.Methods CD138" and CD138+ cells isolated from U266 cell lines by MACS. The β-catenin mRNA expression levels of Wnt1/ β-catenin signaling pathways of U266 multiple myeloma cell lines, human B lymphocytes in peripheral blood, CD 138", and CD138+ cells were detected by RT-PCR method. The shRNAs special for and not for β-catenin were used as interferents for expression of β-catenin produced by CD138-cell. The growth state of CD138- cells in stem cell culture fluid was observed using inverted microscope. Results (1) The expression β-catenin mRNA of U266 cell line was 9.90 times to peripheral blood B lymphocytes of healthy population. CD138+ cells and CD 138- cells in U266 multiple myeloma cell lines accounted for 97.1%,2.9% respectively. (2) The purity quotient of CD 138" cells isolated from U266 cell lines was 71.6%. CD 138" cells growing as a floating sphere-like after cultured for 5 to 7 days in the Stem-cell medium. The level of p-catenin mRNA in CD138" cells were 2.99 times and 2.61 times to U266 cells and CD138+ cells, respectively. (3) The expression levels of β-catenin mRNA and protein in CD 138- cell interferented by the shRNAs special for β-catenin were 14.66% and 48.0% to control CD 138- cells,respectively.The growth of CD 138" cells was inhibited by the shRNAs special for β-catenin.Conclusion(1) The expression of β-catenin mRNA in Wnt1/β-catenin signaling pathway of U266 multiple myeloma cells was higher than B lymphocytes in peripheral blood.(2) The expression of β-catenin mRNA in Wnt1/β-catenin signaling pathway of CD 138-cells isolated from U266 cell lines was higher than U266 and CD 138+ cells.(3) The growth of CD 138-cells and the expression of β-catenin mRNA and protein were affected by β-catenin-shRNA.