Department of Spine Surgery, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, 121000, Liaoning, China.
Respiration Medicine, The First Affiliated Hospital of Jinzhou Medical University, No. 2, Section 5, Renmin Street, Guta District, Jinzhou, 121000, Liaoning, China.
J Orthop Surg Res. 2023 Nov 22;18(1):890. doi: 10.1186/s13018-023-04387-z.
Osteosarcoma is a common malignant bone tumor. Cisplatin (DDP) achieves a high response rate in osteosarcoma. Here we aim to study the dysregulation of long non-coding RNA the growth arrest-specific transcript 5 (GAS5), and its roles in DDP-resistance of osteosarcoma. The expression of mRNA and microRNA in osteosarcoma tissues and osteosarcoma cell lines were detected by quantitative reverse-transcription polymerase chain reaction, and protein expression levels were measured by western blotting assay. Cell Counting Kit-8 and 5-Ethynyl-2'-deoxyuridine were used to measure cell proliferation. Flow cytometer assay was used to evaluate cell apoptosis. The interactions between miR-26b-5p and GAS5 or tumor protein p53-induced nuclear protein 1 (TP53INP1) were verified by dual luciferase reporter along with biotin RNA pull-down assays. GAS5 was identified to be significantly lowly expressed in osteosarcoma samples especially in cisplatin-resistant (DDP-resistant) tissues. GAS5 was also downregulated in DDP-resistant cells. Over-expressed GAS5 prominently increased the sensitivity of osteosarcoma cells to DDP in vitro. Furthermore, over-expression of GAS5 suppressed cell proliferation and facilitated apoptosis of DDP-resistant cells. Mechanistically, GAS5 sponged miR-26b-5p, over-expression of which reversed the effects of GAS5 on cell proliferation and apoptosis of DDP-resistant cells. In addition, miR-26b-5p targeted TP53INP1. TP53INP1 abrogated the functions of miR-26b-5p on cell proliferation and apoptosis in DDP-resistant cells. Taken together, GAS5 enhanced the sensitivity of osteosarcoma cells to DDP via GAS5/miR-26b-5p/TP53INP1 axis. Therefore, GAS5 may be a potential indicator for the management of osteosarcoma.
骨肉瘤是一种常见的恶性骨肿瘤。顺铂（DDP）在骨肉瘤中取得了很高的缓解率。在这里，我们旨在研究长非编码 RNA 生长停滞特异性转录物 5（GAS5）的失调及其在骨肉瘤 DDP 耐药中的作用。通过定量逆转录聚合酶链反应检测骨肉瘤组织和骨肉瘤细胞系中的 mRNA 和 microRNA 表达，通过 Western blot 测定蛋白表达水平。细胞计数试剂盒-8 和 5-乙炔基-2'-脱氧尿苷用于测量细胞增殖。流式细胞仪检测用于评估细胞凋亡。通过双荧光素酶报告基因和生物素 RNA 下拉实验验证 miR-26b-5p 与 GAS5 或肿瘤蛋白 p53 诱导核蛋白 1（TP53INP1）之间的相互作用。
结果表明，GAS5 在骨肉瘤样本中表达明显降低，尤其是在顺铂耐药（DDP 耐药）组织中。DDP 耐药细胞中 GAS5 也下调。过表达 GAS5 显著增加了骨肉瘤细胞对 DDP 的体外敏感性。此外，过表达 GAS5 抑制 DDP 耐药细胞的增殖并促进其凋亡。机制上，GAS5 海绵吸附 miR-26b-5p，过表达 miR-26b-5p 逆转了 GAS5 对 DDP 耐药细胞增殖和凋亡的影响。此外，miR-26b-5p 靶向 TP53INP1。TP53INP1 废除了 miR-26b-5p 在 DDP 耐药细胞中对细胞增殖和凋亡的作用。
总之，GAS5 通过 GAS5/miR-26b-5p/TP53INP1 轴增强骨肉瘤细胞对 DDP 的敏感性。因此，GAS5 可能是骨肉瘤治疗的潜在指标。
