PDF(Pubmed)
Abstract：
The goal of this study was to demonstrate the functions and specific mechanism of long non-coding RNA (lncRNA) GNAS-AS1 in lung adenocarcinoma. Levels of lncRNA GNAS-AS1, microRNA (miR)-433-3p, and Rab3A were assessed by quantitative real-time PCR (qRT-PCR). The target-binding sites of lncRNA GNAS-AS1, miR-433-3p, and Rab3A were predicted and confirmed by bioinformatics tool (StarBase) and a dual-luciferase reporter system. Cell proliferation and apoptosis were checked using MTT and flow cytometry, respectively. Additionally, the levels of apoptosis-related and epithelial-mesenchymal transition (EMT)-associated genes in A549 cells were analyzed by qRT-PCR and western blot. We found that lncRNA GNAS-AS1 was upregulated, miR-433-3p was low-expressed, and Rab3A was overexpressed in lung adenocarcinoma tissues and cell lines. LncRNA GNAS-AS1 interacted with miR-433-3p and negatively regulated miR-433-3p levels. Rab3A was a direct target of miR-433-3p. Downregulation of lncRNA GNAS-AS1 remarkably suppressed cell proliferation, promoted cell apoptosis, decreased B-cell lymphoma-2 (Bcl-2) expression, enhanced the Bcl-2-Associated X (Bax) level, promoted E-cadherin expression, and reduced N-cadherin and Rab3A levels. However, the miR-433-3p inhibitor reversed all these findings. Similarly, the inhibitory effects of miR-433-3p mimic on A549 cells were reversed by the Rab3A-plasmid. In conclusion, lncRNA GNAS-AS1 downregulation suppressed lung adenocarcinoma cell proliferation and EMT through the miR-433-3p/Rab3A axis.
摘要：
这项研究的目的是证明长链非编码RNA（lncRNA）GNAS-AS1在肺腺癌中的功能和特定机制。lncRNAGNAS-AS1，microRNA（miR）-433-3p，和Rab3A通过定量实时PCR(qRT-PCR)评估。lncRNAGNAS-AS1，miR-433-3p，和Rab3A通过生物信息学工具(StarBase)和双荧光素酶报告系统进行预测和确认。用MTT法和流式细胞术检测细胞增殖和凋亡,分别。此外,通过qRT-PCR和Westernblot分析A549细胞中凋亡相关基因和上皮间质转化（EMT）相关基因的水平。我们发现lncRNAGNAS-AS1被上调，miR-433-3p低表达，Rab3A在肺腺癌组织和细胞系中过度表达。LncRNAGNAS-AS1与miR-433-3p相互作用，并负调控miR-433-3p水平。Rab3A是miR-433-3p的直接靶标。lncRNAGNAS-AS1的下调显著抑制了细胞增殖，促进细胞凋亡，B细胞淋巴瘤-2（Bcl-2）表达降低，增强Bcl-2相关X（Bax）水平，促进E-cadherin表达，并降低N-钙粘蛋白和Rab3A水平。然而,miR-433-3p抑制剂逆转了所有这些发现.同样，miR-433-3p模拟物对A549细胞的抑制作用被Rab3A质粒逆转。总之,lncRNAGNAS-AS1下调通过miR-433-3p/Rab3A轴抑制肺腺癌细胞增殖和EMT。
