长链非编码RNA LINC00885在人食管癌细胞中的作用研究

Study on the Role of Long Noncoding RNA LINC00885 in Human Esophageal Cancer Cells

探讨长链非编码RNA LINC00885对人食管癌细胞EC109增殖、迁移与侵袭的影响。构建LINC00885基因过表达质粒(pcDNA3.1-LINC00885)和shRNA敲低质粒(pLKO.1-LINC00885)。分别采用集落形成实验检测EC109细胞增殖能力;划痕实验检测EC109细胞横向迁移能力;Transwell实验检测EC109细胞纵向迁移能力及侵袭能力;流式实验检测LINC00885对于细胞周期的调控;实时定量PCR方法检测LINC00885 mRNA转录水平;Western blot检测BMP7及上皮间充质转化(epithelial-to-mesenchymal transition,EMT)通路相关蛋白(VIMENTIN、β-catenim和ZO-1)表达水平。在过表达LINC00885的EC109细胞中,细胞的增殖能力、迁移能力及侵袭能力均显著增强,BMP7蛋白表达升高;而在敲低表达LINC00885的EC109细胞中,细胞的增殖能力、迁移能力与侵袭能力均显著降低,BMP7蛋白表达也随之降低。另外,在过表达LINC00885的EC109细胞中,EMT通路蛋白VIMENTIN、β-catenim表达水平显著升高,而ZO-1表达水平显著降低。通过探究LINC00885对食管癌细胞增殖、迁移能力的影响,旨在验证LINC00885在食管癌中的功能,以期为临床治疗食管癌奠定理论基础。

To investigate the effect of long non-coding RNA LINC00885 on the proliferation,migration and invasion of human esophageal cancer cell EC109 in this study.The LINC00885 gene overexpression plasmid(pcDNA3.1-LINC00885)and shRNA knockdown plasmid(pLKO.1-LINC00885)were constructed.Colony formation experiment was used to detect the proliferation ability of EC109 cells;the scratch healing experiment was used to detect the lateral migration ability of EC109 cells;the Transwell chamber experiment was used to detect the longitudinal migration ability and invasion ability of EC109 cells;the flow cytometry experiment was used to detect the regulation of LINC00885 on the cell cycle;real-time quantitative PCR method was used to detect the LINC00885 mRNA transcription level;Western blot was used to detect the expression levels of BMP7 and epithelial-to-mesenchymal transition(EMT)pathway related proteins(VIMENTIN,β-catenim and ZO-1).In EC109 cells over⁃expressing LINC00885,the proliferation,migration and invasion capabilities of the cells were significantly enhanced,and the expression of BMP7 protein was increased.While LINC00885 down-expression significantly inhibited the proliferation,migration and invasion capabilities of EC109 cells,and BMP7 protein expression was decreased.In addition,overexpressing LINC00885 in EC109 cells,the expression levels of EMT pathway proteins VIMENTIN andβ-catenim increased significantly,while the ex⁃pression levels of ZO-1 decreased significantly.By exploring the effect of LINC00885 on the proliferation and migration of esopha⁃geal cancer cells,this paper aimed to verify the function of Linc00885 in esophageal cancer,in order to lay a theoretical foundation for clinical treatment of esophageal cancer.