High-throughput sequencing reveals circular RNA hsa_circ_0000592 as a novel player in the carcinogenesis of gastric carcinoma
High-throughput sequencing reveals circular RNA hsa_circ_0000592 as a novel player in the carcinogenesis of gastric carcinoma
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高通量测序揭示环状RNA hsa_circ_0000592是胃癌发生过程中的新参与者
DOI：
10.1042/bsr20181900
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发表时间：
2019-06-28
影响因子：
4
通讯作者：
Zhou, Xinke
中科院分区：
文献类型：
--
作者：
Liang, Min;Liu, Zhaoyu;Zhou, Xinke
背景/目的：胃癌是最常见的恶性肿瘤之一，其复杂的发病机制尚未完全阐明。环状rna （circRNAs）参与各种生物过程和人类疾病。然而，它们的确切功能作用和作用机制在很大程度上仍不清楚。我们之前在人胃上皮细胞的恶性转化过程中发现了非编码rna （ncRNAs）的差异表达。在这项研究中，我们研究了一个显著上调的circRNA （hsa_circ_0000592）在胃癌中的功能作用。方法：采用高通量circRNA测序方法对n -甲基-n ' -硝基-n -亚硝基胍（MNNG）诱导的胃上皮细胞恶性转化（GES-1-T）和正常胃上皮细胞（GES-1-N）进行分析。在不同的胃上皮细胞系中，通过qRT-PCR进一步证实高通量测序结果中上调最多的15个环状rna。通过RNA干扰（RNAi）检测、荧光原位杂交分析（FISH）和生物信息学预测方法研究了最重要的circRNA （hsa_circ_0000592）的功能。结果：与GES-1-N细胞相比，GES-1-T细胞共上调1509个基因，下调3142个基因。与GES-1-N细胞相比，hsa_circ_0000592在GES-1-T细胞以及其他胃癌细胞系中明显上调。hsa_circ_0000592 mRNA的沉默导致细胞增殖减少，细胞周期停滞在G0/G1期，细胞凋亡率增加，细胞迁移减少。此外，FISH显示hsa_circ_0000592主要位于细胞质中，生物信息学分析表明hsa_circ_0000592可能通过吸附多种mirna发挥作用，最明显的是四个保守的mirna，包括miR-139-3p、miR-200、miR-367-3p和miR-33a-3p。结论：本研究首次发现hsa_circ_0000592是一种在mnng诱导的胃癌中起关键作用的新型circRNA。由于hsa_circ_0000592在胃癌细胞中的重要作用，它可能被认为是一种潜在的用于胃癌诊断的生物标志物。我们的发现为环状rna在环境致癌物诱导的胃癌中的功能提供了新的见解。
Background/Aim: Gastric cancer is one of the most common malignant tumors, and its complex pathogenesis has not been fully elucidated. Circular RNAs (circRNAs) are involved in various biological processes and human diseases. However, their exact functional roles and mechanisms of action remain largely unclear. We previously discovered the differential expression of non-coding RNAs (ncRNAs) during the malignant transformation of human gastric epithelial cells. In this study, we investigated the functional roles of a significantly up-regulated circRNA (hsa_circ_0000592) in gastric cancer. Methods: N-methyl-N′-nitro-N-nitrosoguanidine (MNNG)-induced malignant-transformed gastric epithelial cells (GES-1-T) and normal gastric epithelial cells (GES-1-N) were analyzed by high-throughput circRNA sequencing. The top 15 up-regulated circRNAs in high-throughput sequencing results were further confirmed by qRT-PCR in different gastric epithelial cell lines. The function of the most significant circRNA (hsa_circ_0000592) was investigated by using RNA interference (RNAi) assays, fluorescence in situ hybridization analysis (FISH), and bioinformatics prediction methods. Results: A total of 1509 genes were up-regulated and 3142 genes were down-regulated in GES-1-T cells when compared with GES-1-N cells. When compared with GES-1-N cells, hsa_circ_0000592 was obviously up-regulated in GES-1-T cells, as well as in other gastric cancer cell lines. The silencing of hsa_circ_0000592 mRNA led to a decrease in cell proliferation, cell cycle arrest at the G0/G1 phase, an increased rate of apoptosis, and a reduction in cell migration. Furthermore, FISH showed that hsa_circ_0000592 was mainly located in the cytoplasm, and a bioinformatics analysis suggested that hsa_circ_0000592 might function by sponging multiple miRNAs, and most notably four conserved miRNAs, including miR-139-3p, miR-200, miR-367-3p, and miR-33a-3p. Conclusion: This study is the first to identify hsa_circ_0000592 as a novel circRNA with a critical role in MNNG-induced gastric cancer. Due to the essential role of hsa_circ_0000592 in gastric carcinoma cells, it may be considered as a potential biomarker for use in diagnosing gastric carcinoma. Our findings provide a new insight into the function of circRNAs in environmental carcinogen-induced gastric cancer.