Abstract: Objective To investigate the effect of knocking down long non-coding RNA ( lncRNA) LINC01296 on the proliferation capacity and stem-cell characteristics of human osteosarcoma (OS) cells, and to explore the related mechanisms. Methods Human normal osteoblast cellline hFOB1. 19 and human OS cell lines MG63, U2OS and Saos2 were cultured, and the expression level of lncRNA LINC01296 was detected by qRT-PCR. MG63 cells were divided into 4 groups: control group, siRNA negative control group ( negative control siRNA was transfected into cells), LINC01296 siRNA interference group (LINC01296 siRNA was transfected into cells), LINC01296 siRNA + LiCl group ( LINC01296 siRNA was transfected into cells and cultured with 10 mmol / L Wnt / β-catenin pathway activator LiCl for 24 h). The expression level of lncRNA LINC01296 in each group was detected, the survival rate of cells in each group was detected by CCK-8 method, the proliferation of cells was observed by EdU staining, and the spheroid number in each group was detected by tumor cell spheroid formation assay, the expression levels of stem-cell related proteins (Nanog, OCT-4, SOX2) and Wnt / β-catenin pathway-related proteins (β-catenin, c-Myc, Cyclin D1) in cells of each group were detected by Western blotting. Results The relative expressionlevel of lncRNA LINC01296 in the human OS cell lines MG63, U2OS and Saos2 was significantlyhigher than that in the hFOB1. 19 cells (P< 0. 05). The survival rate, the proportion of EdU positive cells, and the number of tumor cell spheres of MG63 cells in the LINC01296 siRNA group wassignificantly decreased when compared with those in the control group (all P< 0. 05), in addition,the relative expression levels of Nanog, OCT-4, SOX2, β-catenin, c-Myc and Cyclin D1 in theLINC01296 siRNA group were significantly down-regulated (P< 0. 05). The survival rate, the proportion of EdU positive cells, and the number of tumor cell spheres of MG63 cells in theLINC01296 siRNA + LiCl group was significantly increased when compared with those in theLINC01296 siRNA group ( all P < 0. 05 ), at the same time, the relative expression levels ofNanog, OCT-4, SOX2, β-catenin, c-Myc and Cyclin D1 in the cells were significantly up-regulated (P< 0. 05). Conclusion Targeted knockout of lncRNA LINC01296 in OS cells can inhibit cellproliferation and reduce osteosarcoma cell stemness, which may be related to inhibiting the activation of Wnt / β-catenin pathway.

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