Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu Province, School of Life Science, Jiangsu Normal University, Xuzhou, P. R. China.
School of Environment Science and Spatial Informatics, China University of Mining and Technology, Xuzhou, P. R. China.
J Cell Biochem. 2018 Feb;119(2):1827-1840. doi: 10.1002/jcb.26344. Epub 2017 Sep 18.
We aim to investigate the interaction between the EZH2 and the long noncoding RNA (lncRNA) SPRY4-IT1. We also explore their respective effects on human lung adenocarcinoma (LA) cell invasion and migration. Both LA and adjacent normal tissues were obtained from 256 LA patients. SPTY4-IT expression and EZH2 mRNA expressions in tissues and cells were detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR). The siRNAs against SPRY4-IT1 and EZH2 were co-transfected into A549 and H1975 cells. The interaction between SPRY4-IT1 and EZH2 was determined using a RNA pull-down assay and a RNA immunoprecipitation (RIP) assay. A Transwell assay and scratch assay were used to evaluate the cell migration and invasion abilities. The expressions of E-cadherin and Vimentin in the epithelial-mesenchymal transition (EMT) and EZH2 protein expression were detected through western blotting. SPRY4-IT1 expression was observed to be significantly lower, while the expression of EZH2 was higher in the LA tissues than in the adjacent normal tissues. Compared with the HBE cell line, expressions of SPRY4-IT1 in each human LA cell line had decreased, with the lowest observed reduction in the A549 cell line, while EZH2 mRNA and protein expression increased in each human LA cell lines. After SPRY4-IT1-siRNA was transfected into A549 and H1975 cells, invasion and migration abilities were enhanced, in addition to a reduction in the expression of E-cadherin, while expressions of Vimentin exhibited an increased rate. Consequently, we find that EZH2 promotes LA cell invasion and metastasis by inhibiting SPRY4-IT1 expression.
我们旨在研究 EZH2 与长链非编码 RNA（lncRNA）SPRY4-IT1 之间的相互作用。我们还探讨了它们各自对人肺腺癌（LA）细胞侵袭和迁移的影响。从 256 名 LA 患者中获得了 LA 组织和相邻正常组织。通过逆转录定量聚合酶链反应（RT-qPCR）检测组织和细胞中 SPRY4-IT1 的表达和 EZH2 mRNA 的表达。将针对 SPRY4-IT1 和 EZH2 的 siRNA 共转染到 A549 和 H1975 细胞中。使用 RNA 下拉测定和 RNA 免疫沉淀（RIP）测定来确定 SPRY4-IT1 和 EZH2 之间的相互作用。通过 Transwell 测定和划痕测定评估细胞迁移和侵袭能力。通过 Western blot 检测上皮-间充质转化（EMT）中 E-cadherin 和 Vimentin 的表达以及 EZH2 蛋白表达。与相邻正常组织相比，LA 组织中 SPRY4-IT1 的表达明显降低，而 EZH2 的表达升高。与 HBE 细胞系相比，每个人类 LA 细胞系中的 SPRY4-IT1 表达均降低，在 A549 细胞系中观察到的降低幅度最大，而每个人类 LA 细胞系中的 EZH2 mRNA 和蛋白表达均增加。将 SPRY4-IT1-siRNA 转染到 A549 和 H1975 细胞中后，侵袭和迁移能力增强，E-cadherin 的表达降低，而 Vimentin 的表达增加。因此，我们发现 EZH2 通过抑制 SPRY4-IT1 的表达促进 LA 细胞的侵袭和转移。
