长链非编码RNA STAG3L5P过表达对口腔鳞状细胞癌细胞增殖和迁移的影响

Effects of long non-coding RNA STAG3L5P overexpression on cell proliferation and migration in oral squamous cell carcinoma

目的:研究长链非编码RNA(long non-coding RNA,lncRNA)STAG3L5P在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)细胞中的表达和定位及其对OSCC细胞增殖及迁移的影响。方法:使用数据库GEPIA2(gene expression profiling interactive analysis 2)分析STAG3L5P在头颈部鳞状细胞癌(head and neck squamous cell carcinoma,HNSC)中的表达;利用数据库UCSC Xena(University of California Santa Cruz Xena)分析STAG3L5P在OSCC中的表达;实时荧光定量PCR(realtime fluorescence quantitative PCR,qPCR)检测STAG3L5P的表达水平;RNA核质分离实验检测其亚细胞定位;细胞计数试剂盒-8(cell counting kit-8,CCK-8)实验和Transwell迁移实验检测STAG3L5P过表达对OSCC细胞增殖和迁移能力的影响;qPCR和Western blot检测STAG3L5P过表达对上皮间充质转化(epithelial-mesenchymal transition,EMT)相关基因的影响;Western blot检测STAG3L5P过表达对PI3K/AKT通路的影响。结果:STAG3L5P在OSCC组织中高表达,且其表达与组织学分级显著相关;STAG3L5P在OSCC细胞的表达水平显著升高,且在细胞质占比明显高于细胞核占比;STAG3L5P过表达组的细胞增殖和迁移能力均显著高于阴性对照组。STAG3L5P过表达导致N-cadherin和Vimentin的mRNA和蛋白表达均上调,E-cadherin蛋白表达下降。STAG3L5P过表达引起p-PI3K和p-AKT表达增多。结论:STAG3L5P在OSCC组织和细胞中高表达,STAG3L5P过表达可以促进OSCC细胞的增殖及迁移能力,这可能与STAG3L5P激活PI3K/AKT通路促进EMT发生有关。

Objective:To investigated the expression and localization of the long non-coding RNA(lncRNA)STAG3L5P in oral squamous cell carcinoma(OSCC)cells and its effects on OSCC cell proliferation and migration.Methods:STAG3L5P expression in HNSC and OSCC was analyzed online using gene expression profiling interactive analysis 2(GEPIA2)and the University of California Santa Cruz Xena(UCSC Xena)database,respectively.STAG3L5P expression in OSCC cell lines was detected using real-time fluorescence quantitative PCR(qPCR).Nuclearcytoplasmic RNA fractionation assays were carried out to pinpoint the location of STAG3L5P.Cell counting kit-8(CCK-8)and Transwell migration assays were used to assess OSCC cell proliferation and migration changes.The effect of STAG3L5P overexpression on epithelial-mesenchymal transition(EMT)related gene expression was detected by qPCR and Western blot.The effect of STAG3L5P overexpression on PI3K/AKT pathway activity was also assessed by Western blot.Results:STAG3L5P was highly expressed in OSCC,and its expression correlated significantly with histological grade.STAG3L5P expression was significantly higher in OSCC cell lines than in normal cells.The level of cytoplasmic STAG3L5P in OSCC cells was significantly higher than that in the nucleus.The proliferation and migration capacity of OSCC cells overexpressing STAG3L5P were significantly enhanced compared to negative control OSCC cells.N-cadherin and vimentin mRNA and protein levels were significantly increased by STAG3L5P overexpression,while E-cadherin protein expression was decreased.Overexpression of STAG3L5P also increased activity of p-PI3K and p-AKT.Conclusions:STAG3L5P is up-regulated in OSCC,and STAG3L5P overexpression can promote OSCC cell proliferation and migration.This effect may be related to activation of the PI3K/AKT pathway,thus promoting EMT.