Pain Treatment Center, The Second Hospital of Tianjin Medical University, Tianjin, China.
Eur Rev Med Pharmacol Sci. 2019 Aug;23(16):6813-6823. doi: 10.26355/eurrev_201908_18720.
The aim of this study was to elucidate the expression pattern and potential function of LINC01116 in regulating the progression of osteosarcoma.
Expression levels of LINC01116 in osteosarcoma tissues (n=52) and adjacent normal tissues (n=52) were detected by quantitative Real-time polymerase chain reaction (qRT-PCR). Survival analysis and univariate analysis were performed in osteosarcoma patients based on the relative expression levels of LINC01116 and clinical data. Overexpression or silence of LINC01116 in osteosarcoma cells was achieved by transfection of plasmid complementary deoxyribonucleic acid (pcDNA)-LINC01116 or si-LINC01116, respectively. Subsequently, the regulatory effects of LINC01116 on cellular behaviors of osteosarcoma cells were examined by cell counting kit-8 (CCK-8), transwell and flow cytometry. Meanwhile, the potential mechanism of LINC01116 in regulating the progression of osteosarcoma was explored by RNA binding protein immunoprecipitation (RIP), chromatin immunoprecipitation (ChIP) and Western blot. Potential target genes in osteosarcoma were searched, and their functions were clarified through a series of rescue experiments.
LINC01116 expression in osteosarcoma tissues was significantly higher than adjacent normal tissues. The expression of LINC01116 was negatively correlated with overall survival, whereas positively correlated with tumor size and clinical grade of osteosarcoma patients. Transfection of pcDNA-LINC01116 significantly enhanced proliferative, migratory and invasive abilities of U2OS cells, shortened G0/G1 phase period, and inhibited cell apoptosis. However, transfection of si-LINC01116 in MG63 cells obtained the opposite trends in the above-mentioned cellular behaviors. Furthermore, RIP assay confirmed the binding of enhancer of zeste homolog 2 (EZH2) to LINC01116. Knockdown of LINC01116 significantly up-regulated the expressions of phosphatase and tensin homolog deleted on chromosome ten (PTEN) and p53. Moreover, EZH2 knockdown could reverse the inhibitory effect of LINC01116 on carcinogenesis of osteosarcoma.
LINC01116 is highly expressed in osteosarcoma. Up-regulated LINC01116 can promote cell proliferation, invasion and cell cycle progression, while inhibiting the apoptosis of osteosarcoma cells. Furthermore, LINC01116 is involved in the development of osteosarcoma by binding to EZH2 to regulate expressions of PTEN and p53.
本研究旨在阐明 LINC01116 在调控骨肉瘤进展中的表达模式和潜在功能。
采用实时定量聚合酶链反应（qRT-PCR）检测 52 例骨肉瘤组织和 52 例相邻正常组织中 LINC01116 的表达水平。根据 LINC01116 的相对表达水平和临床资料，对骨肉瘤患者进行生存分析和单因素分析。通过转染质粒互补脱氧核糖核酸（pcDNA）-LINC01116 或 si-LINC01116 分别实现骨肉瘤细胞中 LINC01116 的过表达或沉默。随后，通过细胞计数试剂盒-8（CCK-8）、Transwell 和流式细胞术检测 LINC01116 对骨肉瘤细胞细胞行为的调节作用。同时，通过 RNA 结合蛋白免疫沉淀（RIP）、染色质免疫沉淀（ChIP）和 Western blot 探讨 LINC01116 调节骨肉瘤进展的潜在机制。搜索骨肉瘤中的潜在靶基因，并通过一系列挽救实验阐明其功能。
骨肉瘤组织中 LINC01116 的表达明显高于相邻正常组织。LINC01116 的表达与骨肉瘤患者的总生存率呈负相关，而与肿瘤大小和临床分级呈正相关。转染 pcDNA-LINC01116 可显著增强 U2OS 细胞的增殖、迁移和侵袭能力，缩短 G0/G1 期，抑制细胞凋亡。然而，在 MG63 细胞中转染 si-LINC01116 则获得了上述细胞行为相反的趋势。此外，RIP 实验证实了增强子的锌指蛋白 2（EZH2）与 LINC01116 的结合。LINC01116 的敲低显著上调了磷酸酶和张力蛋白同源物缺失的第十染色体（PTEN）和 p53 的表达。此外，EZH2 的敲低可逆转 LINC01116 对骨肉瘤发生的抑制作用。
LINC01116 在骨肉瘤中高表达。上调的 LINC01116 可促进骨肉瘤细胞的增殖、侵袭和细胞周期进程，同时抑制骨肉瘤细胞的凋亡。此外，LINC01116 通过与 EZH2 结合调节 PTEN 和 p53 的表达参与骨肉瘤的发生发展。
