Zhou Yiran, Li Ruhong, Yu Haidong, Wang Ruotian, Shen Zhiqiang
Department of Pharmacy, Kunming Medical University.
Yan'an Hospital Affiliated to Kunming Medical University, Kunming, People's Republic of China.
Onco Targets Ther. 2017 Aug 3;10:3893-3905. doi: 10.2147/OTT.S139443. eCollection 2017.
Gastric cancer is one of the most common causes of death worldwide, although its incidence has steadily declined in recent years. There is strong evidence that aberrantly expressed microRNAs (miRNAs) are involved in gastric cancer tumorigenesis. Furthermore, is closely associated with the occurrence and development of gastric cancer, and our predictions suggest that miR-130a, which can promote gastric cancer tumorigenesis, is a potential regulator. In this study, we investigated the expression of and miR-130a in human gastric cancer cell lines by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot (WB) examination and direct interactions between miR-130a and by dual-luciferase reporter assay. We also evaluated the biological roles of miR-130a and in gastric cancer cells by flow cytometry, MTT assay, soft agar colony formation assay, and Transwell tests and confirmed function in vivo, using a tumor xenograft model. Our results demonstrated that expression was significantly decreased at both the gene and protein levels, while miR-130a expression was notably increased, in five human gastric cancer cell lines compared with human gastric epithelial cells. Dual-luciferase reporter assays indicated that was the direct target of miR-130a. Moreover, an inverse regulatory relationship between miR-130a and was verified by qRT-PCR and WB, and overexpression of miR-130a in BGC823 cells enhanced apoptosis and cell proliferation, arrested the cell cycle in G0/G1, and facilitated cell colony formation, invasion, migration, and adhesion, while upregulation of had opposite effects. Finally, the growth and weight of transplanted tumors derived from BGC823 cells in which was knocked down were remarkably reduced. These data indicate that miR-130a is an oncomir targeting and could be developed as a potential prognostic factor and a novel therapeutic target in gastric cancer.
胃癌是全球最常见的死亡原因之一，尽管其发病率近年来呈稳步下降趋势。有强有力的证据表明，异常表达的微小RNA（miRNA）参与了胃癌的肿瘤发生过程。此外， 与胃癌的发生和发展密切相关，我们的预测表明，可促进胃癌肿瘤发生的miR-130a是一种潜在的 调节因子。在本研究中，我们通过定量逆转录聚合酶链反应（qRT-PCR）和蛋白质免疫印迹（WB）检测，研究了 及miR-130a在人胃癌细胞系中的表达情况，并通过双荧光素酶报告基因检测法研究了miR-130a与 的直接相互作用。我们还通过流式细胞术、MTT法、软琼脂集落形成试验和Transwell试验评估了miR-130a和 在胃癌细胞中的生物学作用，并使用肿瘤异种移植模型在体内证实了 的功能。我们的结果表明，与人类胃上皮细胞相比，在五种人胃癌细胞系中， 的基因和蛋白水平表达均显著降低，而miR-130a的表达则显著增加。双荧光素酶报告基因检测表明 是miR-130a的直接靶标。此外，通过qRT-PCR和WB验证了miR-130a与 之间的反向调节关系，在BGC823细胞中过表达miR-130a可增强细胞凋亡和增殖，使细胞周期停滞在G0/G1期，并促进细胞集落形成、侵袭、迁移和黏附，而 的上调则产生相反的效果。最后，敲低 的BGC823细胞衍生的移植瘤的生长和重量显著降低。这些数据表明，miR-130a是靶向 的致癌miRNA，可作为胃癌潜在的预后因素和新型治疗靶点进行开发。
