Luo Yue, Tao Hai, Jin Lin, Xiang Wei, Guo Weichun
Department of Orthopaedics, Renmin Hospital of Wuhan University, Wuhan, P.R. China.
Cancer Biother Radiopharm. 2020 Feb;35(1):58-65. doi: 10.1089/cbr.2019.2885. Epub 2019 Nov 14.
Long non-coding RNAs (lncRNAs) are a new class of noncoding RNAs, which serve as crucial regulators in tumor progression, including osteosarcoma (OS). The objective of this study is to uncover the clinical significance and biological role of lncRNA CDKN2B antisense RNA 1 () in OS. Quantitative real time PCR analysis was used to determine the expression of in OS tissues and cell lines. The associations between expression and clinicopathological characteristics of OS patients were assessed by Chi-squared test. CCK-8, colony formation, flow cytometry and transwell assay were utilized to evaluate the effects of knockdown on cell proliferation, cell cycle, migration and invasion. The protein expression associated with cell cycle and epithelial-mesenchymal transition (EMT) was measured by western blotting. was found to be markedly up-regulated in OS tissues and cell lines. Clinical data further demonstrated highly expressed tended to be strongly positively correlated with tumor size, distant metastasis and TNM stage. Loss-of-function of leaded to inhibited cell proliferation and induced cell cycle G0/G1 phase arrest. In addition, knockdown significantly suppressed OS cells migration and invasion. Mechanistically, knockdown in OS cells suppressed the expression of CDK4 and Cyclin D1, as well as EMT, as demonstrated by elevated levels of epithelial markers (E-cadherin) and downregulation of mesenchymal markers (vimentin and N-cadherin). Taken together, our findings suggest that represents a potential therapeutic target for OS.
长链非编码RNA(lncRNAs)是一类新的非编码RNA,在肿瘤进展中发挥关键调节作用,包括骨肉瘤(OS)。本研究的目的是揭示lncRNA CDKN2B反义RNA 1()在骨肉瘤中的临床意义和生物学作用。采用定量实时PCR分析来确定骨肉瘤组织和细胞系中该lncRNA的表达。通过卡方检验评估该lncRNA表达与骨肉瘤患者临床病理特征之间的关联。利用CCK-8、集落形成、流式细胞术和Transwell实验来评估该lncRNA敲低对细胞增殖、细胞周期、迁移和侵袭的影响。通过蛋白质印迹法检测与细胞周期和上皮-间质转化(EMT)相关的蛋白质表达。结果发现该lncRNA在骨肉瘤组织和细胞系中显著上调。临床数据进一步表明,该lncRNA高表达往往与肿瘤大小、远处转移和TNM分期呈强正相关。该lncRNA功能缺失导致细胞增殖受抑制并诱导细胞周期G0/G1期阻滞。此外,该lncRNA敲低显著抑制骨肉瘤细胞的迁移和侵袭。机制上,骨肉瘤细胞中该lncRNA敲低抑制了CDK4和细胞周期蛋白D1的表达,以及EMT,表现为上皮标志物(E-钙黏蛋白)水平升高和间质标志物(波形蛋白和N-钙黏蛋白)下调。综上所述,我们的研究结果表明该lncRNA是骨肉瘤潜在的治疗靶点。