Chen Xiuhui, Qu Junying
Department of Obstetrics and Gynecology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200021, China,
Department of Obstetrics and Gynecology, 1st Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China.
Onco Targets Ther. 2018 Aug 20;11:4999-5007. doi: 10.2147/OTT.S167053. eCollection 2018.
Long non-coding RNAs nowadays emerge as important biomarkers or potential therapeutic targets discussed in human cancers. Among them, maternally expressed gene 3 (MEG3) is known to be decreased in a variety of malignancies, and this affects tumor cellular proliferation, migration, and invasion.
Quantitative real-time PCR was performed to detect the expression of MEG3 in normal cervical epithelium, cervical intraepithelial neoplasia, and cervical squamous cell carcinoma tissues. Gain-of-function and loss-of-function studies were carried out to determine the effect of MEG3 on cell survival, migration, and invasion, which was evaluated by CCK-8 assay, wound healing assay, and transwell assays. mRNA and protein expression of Rac1 were finally determined by quantitative real-time PCR and immunoblotting, respectively. In addition, rescue experiments were performed by overexpression of Rac1.
The expression of MEG3 was downregulated in cervical intraepithelial neoplasia and squamous cell carcinoma tissues. Forced expression of MEG3 led to reduced abilities of cell survival. Overexpression of MEG3 also inhibited cell migration and invasion in vitro. Cell proliferation marker and EMT markers were changed consistently with the phenotype. In addition, Rac1 was inhibited by MEG3 overexpression at both transcriptional and translational levels. Also, Rac1 could rescue the phenotype caused by long non-coding RNA MEG3. And, it negatively correlated with MEG3 expression in cervical cancer (CC) tissues and cell lines.
Our findings revealed that MEG3 could negatively regulate CC cell survival, migration, and invasion. It might serve as an important target for CC treatment.
长链非编码RNA如今已成为人类癌症中讨论的重要生物标志物或潜在治疗靶点。其中,母源表达基因3(MEG3)在多种恶性肿瘤中表达降低,这影响肿瘤细胞的增殖、迁移和侵袭。
采用定量实时PCR检测MEG3在正常宫颈上皮、宫颈上皮内瘤变和宫颈鳞状细胞癌组织中的表达。进行功能获得和功能缺失研究以确定MEG3对细胞存活、迁移和侵袭的影响,通过CCK-8法、伤口愈合试验和Transwell试验进行评估。最后分别通过定量实时PCR和免疫印迹法测定Rac1的mRNA和蛋白表达。此外,通过Rac1过表达进行挽救实验。
MEG3在宫颈上皮内瘤变和鳞状细胞癌组织中的表达下调。MEG3的强制表达导致细胞存活能力降低。MEG3过表达也抑制体外细胞迁移和侵袭。细胞增殖标志物和EMT标志物与表型一致变化。此外,MEG3过表达在转录和翻译水平均抑制Rac1。而且,Rac1可挽救由长链非编码RNA MEG3引起的表型。并且,它在宫颈癌(CC)组织和细胞系中与MEG3表达呈负相关。
我们的研究结果表明,MEG3可负向调节CC细胞的存活、迁移和侵袭。它可能成为CC治疗的重要靶点。