Feng Qi, Wang Donglai, Guo Peng, Zhang Zibo, Feng Jiangang
Department of Orthopedics, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.
Oncol Lett. 2021 Jun;21(6):444. doi: 10.3892/ol.2021.12705. Epub 2021 Apr 6.
The long non-coding RNA (lncRNA) HOTAIR is an oncogene, that has been reported to be aberrantly expressed in multiple types of malignant tumor tissues. However, its expression and association with synovial sarcoma (SS) remains unclear. The present study aimed to elucidate the expression level of HOTAIR in SS tissues and also identify its role. Reverse transcription-quantitative PCR was used to detect the expression level of HOTAIR and microRNA (miR)-126 in 54 tissue samples from patients with SS, in 10 tissue samples from synovium tissues of normal patients, and in SW982 cells. The protein expression level was measured using western blot analysis and cellular immunofluorescence. Cellular proliferation, invasion and migration were assessed using MTT, Transwell and wound healing assays, respectively. HOTAIR was expressed at high levels in SS tissues. In contrast, miR-126 was expressed at low levels in SS tissues, and was negatively correlated with HOTAIR expression. HOTAIR knockdown in SW982 cells inhibited cellular proliferation , but also significantly increased the ratio of cells in the G/G phase of the cell cycle, and decreased the ratio of cells in the G/S phase. In addition, HOTAIR knockdown inhibited the invasion and migration of the SW982 cells, as observed in the Transwell and wound healing assays. Furthermore, HOTAIR knockdown increased miR-126 expression level and decreased the expression level of stromal cell-derived factor-1 (SDF-1) at the protein level. On the other hand, while miR-126-mimic decreased the protein expression level of SDF-1, miR-126-inhibitor increased its expression level in SW982 cells. Notably, HOTAIR knockdown or SDF-1 knockout significantly decreased the protein expression levels of CDK1, CDK2, cyclin D1, MMP-9, vimentin and N-cadherin, and significantly increased the protein expression levels of p21, p53 and E-cadherin in SW982 cells. HOTAIR was highly expressed in SS tissues, wherein it could promote the proliferation, invasion and migration of SS cells by increasing the expression of SDF-1 via miR-126 inhibition.
长链非编码RNA（lncRNA）HOTAIR是一种癌基因，据报道在多种恶性肿瘤组织中异常表达。然而，其在滑膜肉瘤（SS）中的表达及相关性仍不清楚。本研究旨在阐明HOTAIR在SS组织中的表达水平，并确定其作用。采用逆转录定量PCR检测54例SS患者组织样本、10例正常患者滑膜组织样本及SW982细胞中HOTAIR和微小RNA（miR）-126的表达水平。采用蛋白质印迹分析和细胞免疫荧光法检测蛋白质表达水平。分别采用MTT法、Transwell法和伤口愈合试验评估细胞增殖、侵袭和迁移能力。HOTAIR在SS组织中高表达。相反，miR-126在SS组织中低表达，且与HOTAIR表达呈负相关。SW982细胞中HOTAIR基因敲低抑制细胞增殖，但也显著增加细胞周期G/G期细胞比例，降低G/S期细胞比例。此外，如Transwell试验和伤口愈合试验所示，HOTAIR基因敲低抑制SW982细胞的侵袭和迁移。此外，HOTAIR基因敲低增加miR-126表达水平，并在蛋白质水平降低基质细胞衍生因子-1（SDF-1）的表达水平。另一方面，miR-126模拟物降低SW982细胞中SDF-1的蛋白质表达水平，而miR-126抑制剂则增加其表达水平。值得注意的是，HOTAIR基因敲低或SDF-1基因敲除显著降低SW982细胞中CDK1、CDK2、细胞周期蛋白D1、MMP-9、波形蛋白和N-钙黏蛋白蛋白表达水平，并显著增加p21、p53和E-钙黏蛋白蛋白表达水平。HOTAIR在SS组织中高表达，通过抑制miR-126增加SDF-1表达，从而促进SS细胞增殖、侵袭和迁移。
