Lu Gongbiao, Du Lin, Guo Yi, Xing Baohua, Lu Jishou, Wei Yanchun
Department of Spine Surgery, Jining No. 1 People's Hospital, Jining, Shandong 272111, P.R. China.
College of Nursing, Jining Medical University, Jining, Shandong 272013, P.R. China.
Exp Ther Med. 2018 Feb;15(2):1934-1940. doi: 10.3892/etm.2017.5649. Epub 2017 Dec 15.
The aim of the current study was to investigate the expression and role of microRNA (miR)-1271 in the pathogenesis of osteosarcoma, and the associated underlying mechanisms. Tissue samples from 45 patients with osteosarcoma were collected, while the 143B, MG-63 and U-2 OS osteosarcoma cell lines were also cultured. The expression levels of miR-1271 in the tissues and cells were detected with reverse transcription-quantitative polymerase chain reaction, and 143B osteosarcoma cells were subjected to miR-1271 manipulation. In addition, the cell proliferation, cell cycle progression, and migration and invasion abilities were assessed by Cell Counting Kit-8 assay, flow cytometry and Transwell chamber assay, respectively. Tissue inhibitor of metalloproteinases 2 (TIMP2) expression level was also detected with western blot analysis. Dual-luciferase reporter assay was performed to investigate the interaction between miR-1271 and TIMP2. The results revealed that miR-1271 expression was significantly elevated in the osteosarcoma tissue and was closely correlated with the clinical TNM staging. The expression levels of miR-1271 were also upregulated in the osteosarcoma cells, with the highest expression observed in 143B cells. Inhibition of miR-1271 significantly inhibited the cell proliferation, G1/S phase transition, and the migration and invasion abilities of 143B cells, while it also resulted in upregulated TIMP2 expression in these cells. Furthermore, overexpression of TIMP2 significantly inhibited the cell proliferation, G1/S phase transition, and migration and invasion abilities of 143B cells. Dual-luciferase reporter assay demonstrated that miR-1271 targeted on the 3'-untranslated region of TIMP2 mRNA. In conclusion, the expression levels of miR-1271 were significantly elevated in osteosarcoma tissues and cells. miR-1271 downregulated the expression of TIMP2 to promote the proliferation and enhance the migration and invasion abilities of 143B osteosarcoma cells, functioning as an oncogene.
本研究旨在探讨微小RNA（miR）-1271在骨肉瘤发病机制中的表达及作用，以及相关潜在机制。收集45例骨肉瘤患者的组织样本，并培养143B、MG-63和U-2 OS骨肉瘤细胞系。采用逆转录-定量聚合酶链反应检测组织和细胞中miR-1271的表达水平，并对143B骨肉瘤细胞进行miR-1271调控。此外，分别通过细胞计数试剂盒-8法、流式细胞术和Transwell小室法评估细胞增殖、细胞周期进程以及迁移和侵袭能力。采用蛋白质免疫印迹分析检测金属蛋白酶组织抑制剂2（TIMP2）的表达水平。进行双荧光素酶报告基因检测以研究miR-1271与TIMP2之间的相互作用。结果显示，miR-1271在骨肉瘤组织中的表达显著升高，且与临床TNM分期密切相关。miR-1271在骨肉瘤细胞中的表达水平也上调，在143B细胞中表达最高。抑制miR-1271可显著抑制143B细胞的增殖、G1/S期转换以及迁移和侵袭能力，同时还导致这些细胞中TIMP2表达上调。此外，TIMP2过表达显著抑制143B细胞的增殖、G1/S期转换以及迁移和侵袭能力。双荧光素酶报告基因检测表明，miR-1271靶向TIMP2 mRNA的3'-非翻译区。综上所述，miR-1271在骨肉瘤组织和细胞中的表达水平显著升高。miR-1271通过下调TIMP2的表达来促进143B骨肉瘤细胞的增殖并增强其迁移和侵袭能力，发挥癌基因的作用。
