Huang Yingying, Du Xiaoxiao, Chen Xiangliu, Chen Chuanzhi, Wang Haiyong, Yang Yan, Teng Lisong
Department of oncological surgery, The First Affiliated Hospital, College of Medicine, Zhejiang University.
Cancer Institute (Key Laboratory for Cancer Intervention and Prevention, China National Ministry of Education, Zhejiang Provincial Key Laboratory of Molecular Biology in Medical Sciences), The Second Affiliated Hospital, Zhejiang University School of Medicine, China.
J Cancer. 2021 Jul 6;12(18):5394-5403. doi: 10.7150/jca.59747. eCollection 2021.
Gastric cancer (GC) is a type of highly malignant cancer. Although the diagnostic and therapeutic methods are innovating, the outcome of GC patients is still poor. Therefore, our research was carried out to explore potential molecular mechanism in the diagnosis of GC. Bioinformatics analyses were used to obtain microRNA and target mRNA of interest. The expression level of miR-301a-5p and Scinderin (SCIN) mRNA were detected by quantitative real-time PCR (qRT-PCR). Western blot assay was used to investigate SCIN protein level. Cell Counting Kit-8 assay (CCK-8) and colony formation assay were used to investigate cell proliferation ability. Transwell assay was employed to examine cell motility. The interaction between miR-301a-5p and SCIN mRNA was verified by dual-luciferase reporter assay. The qRT-PCR analysis revealed that the expression of miR-301a-5p was higher in gastric cancer tissues than para-cancer tissues (P<0.05). Cox regression analysis showed upregulated miR-301a-5p was associated with larger tumor size (P=0.036) and more advanced TNM stage (P=0.048). The Kaplan-Meier analysis showed a correlation between increased miR-301a-5p expression and shorter overall survival (OS)(P=0.018). By using bioinformatic analysis, SCIN was predicted as one of the targets of miR-301a-5p. Overexpressing miR-301a-5p promoted proliferation and motility of GC cells while knockdown of SCIN exhibited the same performance. Further, we verified the alteration of miR-301a-5p and SCIN expression level could affect the epithelial-mesenchymal transition (EMT) progression on GC cells via STAT3 and NF-κB signaling. Highly expressed miR-301a-5p was associated with aggressiveness of GC. Upregulation of miR-301a-5p promoted malignant phenotype of GC by targeting SCIN. The present results indicated miR-301a-5p might be a promising molecule in the prognosis of GC.
胃癌（GC）是一种高度恶性的癌症。尽管诊断和治疗方法不断创新，但GC患者的预后仍然很差。因此，我们开展了这项研究以探索GC诊断中的潜在分子机制。利用生物信息学分析来获取感兴趣的微小RNA和靶标信使核糖核酸（mRNA）。通过定量实时聚合酶链反应（qRT-PCR）检测miR-301a-5p和肌动蛋白切割蛋白（SCIN）mRNA的表达水平。采用蛋白质免疫印迹法检测SCIN蛋白水平。使用细胞计数试剂盒-8法（CCK-8）和集落形成试验来研究细胞增殖能力。采用Transwell试验检测细胞迁移能力。通过双荧光素酶报告基因试验验证miR-301a-5p与SCIN mRNA之间的相互作用。qRT-PCR分析显示，miR-301a-5p在胃癌组织中的表达高于癌旁组织（P<0.05）。Cox回归分析表明，miR-301a-5p上调与肿瘤体积较大（P=0.036）和TNM分期更晚（P=0.048）相关。Kaplan-Meier分析显示，miR-301a-5p表达增加与总生存期（OS）缩短相关（P=0.018）。通过生物信息学分析，预测SCIN是miR-301a-5p的靶标之一。过表达miR-301a-5p可促进GC细胞的增殖和迁移，而敲低SCIN也表现出相同的效果。此外，我们证实miR-301a-5p和SCIN表达水平的改变可通过信号转导和转录激活因子3（STAT3）和核因子κB（NF-κB）信号通路影响GC细胞的上皮-间质转化（EMT）进程。高表达的miR-301a-5p与GC的侵袭性相关。miR-301a-5p上调通过靶向SCIN促进GC的恶性表型。目前的结果表明，miR-301a-5p可能是GC预后中有前景的分子。
