Li Peifei, Chen Huilin, Chen Shengcan, Mo Xiaoyan, Li Tianwen, Xiao Bingxiu, Yu Rui, Guo Junming
Department of Biochemistry and Molecular Biology, Zhejiang Key Laboratory of Pathophysiology, Ningbo University School of Medicine, Ningbo 315211, China.
Department of Human Body Function, Ningbo College of Health Sciences, Ningbo 315010, China.
Br J Cancer. 2017 Feb 28;116(5):626-633. doi: 10.1038/bjc.2016.451. Epub 2017 Jan 12.
Circular RNAs (circRNAs) are a class of non-coding RNAs broadly expressed in cells of various species. Their role in cancers, especially in gastric cancer, is poorly understood.
Circular RNA 0000096 (hsa_circ_0000096) levels in 101 paired gastric cancer tissues and adjacent non-tumorous tissues from patients with gastric cancer were detected by real-time quantitative reverse transcription-polymerase chain reaction. A receiver operating characteristic curve was generated to evaluate the diagnostic value of hsa_circ_0000096. RNA interference was used to manipulate the expression of hsa_circ_0000096. Its biological effects were evaluated by flow cytometry, real-time cell analysis, a wound scratch assay, western blot analysis and xenograft models.
Hsa_circ_0000096 was found to be significantly downregulated in gastric cancer tissues and gastric cancer cell lines compared with paired adjacent non-tumorous tissues and normal gastric epithelial cells (P<0.001). Moreover, knockdown of hsa_circ_0000096 significantly inhibited cell proliferation and migration in vitro and in vivo. The results of both immunohistochemical and western blot analyses showed that the protein levels of cyclin D1, cyclin-dependent kinase 6 (CDK6), matrix metalloproteinase-2 and MMP-9 were significantly reduced in vitro and in vivo. A gastric cancer xenograft nude mouse model indicated that Ki67 and VEGF were reduced in a dose-dependent manner following knockdown of hsa_circ_0000096. However, the expression of E-cadherin increased.
Hsa_circ_0000096 may be used as a potential novel biomarker for gastric cancer. It affects gastric cancer cell growth and migration by regulating cyclin D1, CDK6, MMP-2 and MMP-9.
环状RNA（circRNAs）是一类在各种物种的细胞中广泛表达的非编码RNA。它们在癌症，尤其是胃癌中的作用尚不清楚。
采用实时定量逆转录-聚合酶链反应检测101例胃癌患者配对的胃癌组织及癌旁非肿瘤组织中环状RNA 0000096（hsa_circ_0000096）的水平。绘制受试者工作特征曲线以评估hsa_circ_0000096的诊断价值。采用RNA干扰技术调控hsa_circ_0000096的表达。通过流式细胞术、实时细胞分析、划痕实验、蛋白质印迹分析和异种移植模型评估其生物学效应。
与配对的癌旁非肿瘤组织和正常胃上皮细胞相比，hsa_circ_0000096在胃癌组织和胃癌细胞系中显著下调（P<0.001）。此外，敲低hsa_circ_0000096可显著抑制体外和体内细胞增殖和迁移。免疫组织化学和蛋白质印迹分析结果均显示，细胞周期蛋白D1、细胞周期蛋白依赖性激酶6（CDK6）、基质金属蛋白酶-2和基质金属蛋白酶-9的蛋白水平在体外和体内均显著降低。胃癌异种移植裸鼠模型表明，敲低hsa_circ_0000096后，Ki67和血管内皮生长因子呈剂量依赖性降低。然而，E-钙黏蛋白的表达增加。
hsa_circ_0000096可能作为胃癌潜在的新型生物标志物。它通过调节细胞周期蛋白D1、CDK6、基质金属蛋白酶-2和基质金属蛋白酶-9影响胃癌细胞的生长和迁移。
