Zhao Juan, Zeng Xue-Bin, Zhang Hong-Yan, Xiang Jie-Wei, Liu Yu-Song
Department of Clinical Laboratory, East Hospital of Sichuan People's Hospital, Sichuan Academy of Medical Sciences, No. 585 Honghe North Road, Chengdu, Sichuan, 610101, China.
Department of Outpatient, East Hospital of Sichuan People's Hospital, Sichuan Academy of Medical Sciences, Chengdu, Sichuan, 610101, China.
Open Med (Wars). 2020 Sep 30;15(1):921-931. doi: 10.1515/med-2020-0175. eCollection 2020.
Long non-coding RNA forkhead box D2 adjacent opposite strand RNA 1 (FOXD2-AS1) has emerged as a potential oncogene in several tumors. However, its biological function and potential regulatory mechanism in glioma have not been fully investigated to date. In the present study, RT-qPCR was conducted to detect the levels of FOXD2-AS1 and microRNA (miR)-506-5p, and western blot assays were performed to measure the expression of CDK2, cyclinE1, P21, matrix metalloproteinase (MMP)7, MMP9, N-cadherin, E-cadherin and vimentin in glioma cells. A luciferase reporter assay was performed to verify the direct targeting of miR-506-5p by FOXD2-AS1. Subsequently, cell viability was analyzed using the CCK-8 assay. Cell migration and invasion were analyzed using Transwell and wound healing assays, respectively. The results demonstrated that FOXD2-AS1 was significantly overexpressed in glioma cells, particularly in U251 cells. Knockdown of FOXD2-AS1 in glioma cells significantly inhibited cell proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) and regulated the expression of CDK2, cyclinE1, P21, MMP7 and MMP9. Next, a possible mechanism for these results was explored, and it was observed that FOXD2-AS1 binds to and negatively regulates miR-506-5p, which is known to be a tumor-suppressor gene in certain human cancer types. Furthermore, overexpression of miR-506-5p significantly inhibited cell proliferation, migration, invasion and EMT, and these effects could be reversed by transfecting FOXD2-AS1 into the cells. In conclusion, our data suggested that FOXD2-AS1 contributed to glioma proliferation, metastasis and EMT via competitively binding to miR-506-5p. FOXD2-AS1 may be a promising target for therapy in patients with glioma.
长链非编码RNA叉头框D2相邻反义链RNA1（FOXD2-AS1）已成为多种肿瘤中的潜在癌基因。然而，其在胶质瘤中的生物学功能和潜在调控机制迄今尚未得到充分研究。在本研究中，进行逆转录定量聚合酶链反应（RT-qPCR）以检测FOXD2-AS1和微小RNA（miR）-506-5p的水平，并进行蛋白质印迹分析以检测胶质瘤细胞中细胞周期蛋白依赖性激酶2（CDK2）、细胞周期蛋白E1、P21、基质金属蛋白酶（MMP）7、MMP9、N-钙黏蛋白、E-钙黏蛋白和波形蛋白的表达。进行荧光素酶报告基因检测以验证FOXD2-AS1对miR-506-5p的直接靶向作用。随后，使用细胞计数试剂盒-8（CCK-8）检测分析细胞活力。分别使用Transwell和划痕愈合实验分析细胞迁移和侵袭能力。结果表明，FOXD2-AS1在胶质瘤细胞中显著过表达，尤其是在U251细胞中。敲低胶质瘤细胞中的FOXD2-AS1可显著抑制细胞增殖、迁移、侵袭和上皮-间质转化（EMT），并调节CDK2、细胞周期蛋白E1、P21、MMP7和MMP9的表达。接下来，探究了这些结果的可能机制，观察到FOXD2-AS1与miR-506-5p结合并对其起负调控作用，已知miR-506-5p在某些人类癌症类型中是一种肿瘤抑制基因。此外，miR-506-5p的过表达显著抑制细胞增殖、迁移、侵袭和EMT，而将FOXD2-AS1转染到细胞中可逆转这些作用。总之，我们的数据表明，FOXD2-AS1通过竞争性结合miR-506-5p促进胶质瘤的增殖、转移和EMT。FOXD2-AS1可能是胶质瘤患者治疗的一个有前景的靶点。
