医学分子生物学杂志2025，Vol.22Issue(2)：166-172,7.DOI:10.3870/j.issn.1672-8009.2025.02.010
敲低lncRNA LINC01296通过Wnt/β-catenin通路抑制骨肉瘤细胞增殖能力及干性特征
Knockdown of lncRNA LINC01296 Inhibited Proliferation and Stem-cell Characteristics of Osteosarcoma Cells through Wnt/β-catenin Pathway
摘要
Abstract
Objective To investigate the effect of knocking down long non-coding RNA(ln-cRNA)LINC01296 on the proliferation capacity and stem-cell characteristics of human osteosarco-ma(OS)cells,and to explore the related mechanisms.Methods Human normal osteoblast cell line hFOB1.19 and human OS cell lines MG63,U2OS and Saos2 were cultured,and the expres-sion 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 de-tected 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,Cyc-lin D1)in cells of each group were detected by Western blotting.Results The relative expression level of lncRNA LINC01296 in the human OS cell lines MG63,U2OS and Saos2 was significantly higher than that in the hFOB1.19 cells(P<0.05).The survival rate,the proportion of EdU posi-tive cells,and the number of tumor cell spheres of MG63 cells in the LINC01296 siRNA group was significantly 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 the LINC01296 siRNA group were significantly down-regulated(P<0.05).The survival rate,the pro-portion of EdU positive cells,and the number of tumor cell spheres of MG63 cells in the LINC01296 siRNA+LiCl group was significantly increased when compared with those in the LINC01296 siRNA group(all P<0.05),at the same time,the relative expression levels of Nanog,OCT-4,SOX2,β-catenin,c-Myc and Cyclin D1 in the cells were significantly up-regula-ted(P<0.05).Conclusion Targeted knockout of lncRNA LINC01296 in OS cells can inhibit cell proliferation and reduce osteosarcoma cell stemness,which may be related to inhibiting the activa-tion of Wnt/β-catenin pathway.关键词
骨肉瘤/长链非编码RNA LINC01296/增殖/肿瘤干性/Wnt/β-catenin通路Key words
osteosarcoma/long non-coding RNA LINC01296/proliferation/stem cell/Wnt/β-catenin pathway分类
临床医学引用本文复制引用
刘广飞,郭志远,王艳花,卢守亮,郭磊,程才..敲低lncRNA LINC01296通过Wnt/β-catenin通路抑制骨肉瘤细胞增殖能力及干性特征[J].医学分子生物学杂志,2025,22(2):166-172,7.基金项目
河北省卫生健康委科研基金(No.20220381) This work was supported by a grant from the Research Fund of Hebei Provincial Health Commission(No.20220381) （No.20220381）
