长链非编码 RNA TUG1 通过调控 miR-212-3p/FOXA1 轴促进骨肉瘤细胞增殖并抑制凋亡。

LncRNA TUG1 promotes cell proliferation and suppresses apoptosis in osteosarcoma by regulating miR-212-3p/FOXA1 axis.

机构信息

Department of Orthopedics, The Second Affiliated Hospital of Guangzhou Medical University, Guangdong, 510260, China.

出版信息

Biomed Pharmacother. 2018 Jan;97:1645-1653. doi: 10.1016/j.biopha.2017.12.004. Epub 2017 Dec 8.

DOI:10.1016/j.biopha.2017.12.004

PMID:29793327

Abstract

BACKGROUND

LncRNA taurine upregulated gene 1 (TUG1) was reported to act as a possible oncogene in osteosarcoma (OS) development. However, the underlying molecular basis of TUG1 involved in the progression of OS remains to be thoroughly investigated.

METHODS

The expressions of TUG1 and miR-212-3p in OS tissues and cells were examined by RT-qPCR. Cell proliferation, apoptosis, caspase-3 activity, protein levels of BCL2, Bax, and forkhead box A1 (FOXA1) were detected by colony formation assay, MTT assay, flow cytometry analysis, caspase-3 activity assay, and western blot. Luciferase reporter assay, RNA immunoprecipitation (RIP) assay, and RT-qPCR were used to explore the interaction between TUG1, FOXA1 and miR-212-3p. Tumor xenograft mouse model was used to confirm the biological role of TUG in OS in vivo.

RESULTS

Elevated TUG1 and FOXA1 expression and reduced miR-212-3p expression were observed in OS tissues and cells. TUG1 knockdown suppressed OS cell proliferation and promoted apoptosis. TUG1 functioned as a ceRNA of miR-212-3p and suppressed miR-212-3p expression. miR-212-3p inhibition reversed the effect of TUG1 knockdown on OS cell proliferation and apoptosis. In addition, FOXA1 was identified as a target of miR-212-3p and TUG1 functioned as a ceRNA to upregulate FOXA1 by sponging miR-212-3p in OS cells. FOXA1 up-regulation abolished the effects of miR-212-3p on OS cell proliferation and apoptosis.

CONCLUSION

TUG1 promoted OS cell proliferation and suppressed apoptosis by regulating the miR-212-3p/FOXA1 axis. Therefore, TUG1/miR-212-3p/FOXA1 axis may be a promising therapeutic target in OS treatment.

摘要

背景

长链非编码 RNA 牛磺酸上调基因 1（TUG1）被报道在骨肉瘤（OS）发展中作为一种可能的癌基因发挥作用。然而，TUG1 参与 OS 进展的潜在分子基础仍需深入研究。

方法

采用 RT-qPCR 检测 OS 组织和细胞中 TUG1 和 miR-212-3p 的表达。采用集落形成实验、MTT 实验、流式细胞术分析、caspase-3 活性测定、western blot 检测细胞增殖、细胞凋亡、caspase-3 活性、BCL2、Bax 和叉头框蛋白 A1（FOXA1）蛋白水平。采用荧光素酶报告基因实验、RNA 免疫沉淀（RIP）实验和 RT-qPCR 探讨 TUG1、FOXA1 和 miR-212-3p 之间的相互作用。采用肿瘤异种移植小鼠模型体内证实 TUG1 在 OS 中的生物学作用。

结果

OS 组织和细胞中 TUG1 和 FOXA1 表达升高，miR-212-3p 表达降低。TUG1 敲低抑制 OS 细胞增殖并促进细胞凋亡。TUG1 作为 miR-212-3p 的 ceRNA 发挥作用，抑制 miR-212-3p 的表达。miR-212-3p 抑制逆转了 TUG1 敲低对 OS 细胞增殖和凋亡的影响。此外，FOXA1 被鉴定为 miR-212-3p 的靶基因，TUG1 通过海绵吸附 miR-212-3p 在 OS 细胞中作为 ceRNA 上调 FOXA1。FOXA1 的上调消除了 miR-212-3p 对 OS 细胞增殖和凋亡的影响。