Regulatory mechanism of casein kinase 1α/chromobox homolog 4 axis on the proliferation and metastasis of osteosarcoma cells
Objective To explore regulatory mechanism of casein kinase 1α(CK1 α)/chromobox homolog 4(CBX4)axis on the proliferation and metastasis of osteosarcoma cells.Methods Transfection of CK1 α over-expression vector(over-expression group)and negative control vector(control group)into U-2OS osteosarcoma cell was used by Lipofectamine™ 3000.The transfection effect was detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and Western blotting.Cell via-bility was measured by methyl thiazolyl tetrazolium(MTT)assay.Cell clone ability was detected by cell clone formation test.Cell apoptosis was detected by Hoechst staining.Cell invasion ability was detected by Transwell method.The expression of B-lymphomatoma-2(bcl-2),bcl-2 related protein X(bax),matrix metalloproteinase(MMP)-2,MMP-9,E-cadherin,Vimentin,Wnt3a,β-catenin and p-glycogen synthase kinase-3β(p-GSK-3 β)was detected by Western blotting.Inter group differences was adopted by independ-ent-samples T-test.Results The expression of CK1α mRNA and protein in over-expression group was higher than that in control group(1.87±0.04 vs.1.00±0.06,2.13±0.12 vs.0.32±0.02,t=6.346,6.893,P<0.05).Cell apoptotic rate,the expression of Wnt3a,β-catenin,bcl-2,MMP-2,MMP-9 and Vimentin protein in over-expression group was higher than that in control group[(36.98±3.62)%vs.(4.31±0.30)%,1.02±0.06 vs.0.22±0.01,0.96±0.07 vs.0.32±0.02,0.88±0.06 vs.0.33±0.02,0.92±0.07 vs.0.28±0.01,0.87±0.05 vs.0.28±0.02,0.97±0.07 vs.0.35±0.02,t=6.671,6.879,5.967,6.345,5.631,6.124,5.983,P<0.05].Cell viability,cell clone number,cell invasion number,the expression of CBX4,p-GSK-3β,bax and E-cadherin protein in over-expression group was lower than that in control group(0.35±0.01 vs.0.58±0.03,32.61±2.68 vs.184.49±12.42,33.74±2.62 vs.161.32±10.58,1.98±0.11 vs.0.30±0.02,0.13±0.01 vs.0.72±0.05,0.14±0.01 vs.0.71±0.05,0.11±0.00 vs.0.63±0.04,t=5.897,6.894,6.692,6.672,5.872,5.652,6.126,P<0.05).Conclusion Over-regulation expression of CK1α might inhibit the proliferation and metastasis of U-2OS osteosarcoma cell,which related to inhibition of Wnt/β-catenin signaling pathway.