The Research Of The Biological Characteristics Of MG-63 Cells After RIPK4 Gene Inhibited By SiRNA

Objective:To investigate the expression and significance of RIPK4 protein in human osteosarcomatous tissue and MG-63 cells;the effect of siRNA-induced down –regulation of expression of Receptor-interacting-protein kinase 4(RIPK4) gene on proliferation and apoptosis of human osteosarcoma MG63 cells andexplore its possible mechanism.Methods:Imunohistochemistry and RT-PCR was performed to detect the expression of RIPK4 protein in paraffin speciments of osteosarcoma patients and RIPK4 mRNA in MG-63 cells, The osteosarcoma MG63 cells were transfected with siRNA targeting RIPK4 gene(RIPK4 siRNA),then the proliferation of human osteosarcoma MG63 cells was by CCK-8 method, the apoptosis was detected by flow cytometry(FCM),and the mRNA and protein expression levels of RIPK4,β-catenin,cyclinD1,Bcl-2,LC3 and Beclin1 were detected by real-time PCR and Western blotting,respectively.Results:The positive expression of RIPK4 protein and mRNA in osteosarcoma and MG-63 cells, The differences were statistically significant(p<0.05);The proliferation ability of MG-63 cells after transfected with RIPK4 siRNA was lower that of MG-63 cells transfected with the control siRNA(as a negative control) or without any transfection(as a blank control)(both p<0.05).The mRNA and protein expression levels of RIPK4,β-catenin and cyclinD1 in RIPK4 siRNA transfection group were lower than those of the negative control and the blank control group(all p<0.05);The expression levels of anti-apoptosis is factor Bcl-2 mRNA and protein in MG-63 cells after transfection with RIPK4 siRNA were down-regulated(p<0.05). The expression levels of autophagy-relate LC3 and Beclin1 mRNAs and Beclin1 protein in MG-63 cells after transfection with RIPK4 siRNA were up-regulated(p<0.05).Conclusion:The expression of RIPK4 protein and gene in osteosarcoma tissues, RIPK4 may be involved in the progress of etithelial osteosarcoma; Silencing the expression of RIPK4 gene can inhibit the proliferation of MG-63 cells. This effect may be related to the inhibition of Wnt/β-catenin signaling pathway;Silencing of RIPK4 gene may promote the apoptosis and autophagy of osteosarcoma MG-63 cells.