MiR-769-5p functions as an oncogene by down-regulating RYBP expression in gastric cancer
MiR-769-5p functions as an oncogene by down-regulating RYBP expression in gastric cancer
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DOI：
10.26355/eurrev_202006_21657
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发表时间：
2020-01-01
影响因子：
3.3
通讯作者：
Yao, J.
中科院分区：
文献类型：
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作者：
Luan, P-B;Jia, X-Z;Yao, J.
目的：本研究旨在检测微小核糖核酸（miR）-769 - 5p在胃癌（GC）组织和细胞中的相对表达水平，并探讨miR - 769 - 5p在胃癌中的临床意义、生物学功能及作用机制。
患者与方法：采用实时定量聚合酶链反应（qRT - PCR）检测62例胃癌组织及癌旁组织中miR - 769 - 5p的相对表达水平。通过卡方检验分析miR - 769 - 5p表达与胃癌患者临床病理特征之间的相关性。此外，采用qRT - PCR检测胃癌细胞中miR - 769 - 5p的相对表达水平以及si - miR - 769 - 5p的干扰效率，并通过体外实验[噻唑蓝（MTT）、流式细胞术、5 - 乙炔基 - 2'-脱氧尿苷（EdU）]研究miR - 769 - 5p的生物学功能。接着，通过裸鼠致瘤实验研究miR - 769 - 5p对胃癌细胞体内致瘤性的影响。此外，通过生物信息学预测miR - 769 - 5p的下游靶基因。最后，采用qRT - PCR和蛋白质印迹法筛选下游靶基因。
结果：在62例胃癌组织中，48例miR - 769 - 5p表达上调。将miR - 769 - 5p分为高表达组和低表达组。卡方分析显示，miR - 769 - 5p高表达与胃癌患者的肿瘤 - 淋巴结 - 转移（TNM）分期（p = 0.005）、淋巴结转移（p = 0.010）以及浸润深度（p = 0.011）呈正相关。qRT - PCR结果表明，miR - 769 - 5p在胃癌细胞中表达上调。体外实验（MTT、流式细胞术、EdU）结果显示，干扰miR - 769 - 5p表达后，胃癌细胞的增殖能力下降，凋亡增加。此外，体内实验结果表明，干扰miR - 769 - 5p表达后，胃癌细胞的致瘤能力下降。最后，qRT - PCR和蛋白质印迹法结果显示，RING1和YY1结合蛋白（RYBP）的表达受miR - 769 - 5p调控。
结论：miR - 769 - 5p在胃癌中表达上调，且与胃癌患者的TNM分期呈正相关。通过调控RYBP的表达，促进胃癌细胞增殖，抑制凋亡。
OBJECTIVE: The purpose of this study was to detect the relative expression level of micro-ribonucleic acid (miR)-769-5p in gastric cancer (GC) tissues and cells, and to investigate the clinical significance, biological function, and mechanism of miR-769-5p in GC.PATIENTS AND METHODS: The relative expression level in 62 cases of GC tissues and paracancerous tissues was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between miR-769-5p expression and clinicopathological characteristics of GC patients was analyzed by Chi-square test. Besides, the relative expression level of miR-769-5p in GC cells and the interference efficiency of si-miR-769-5p were detected by qRT-PCR, and the biological function of miR-769-5p was studied by in vitro experiments [Thiazolyl Blue Tetrazolium Bromide (MTT), flow cytometry, 5-ethynyl-2'-deoxyuridine (EdU)] Next, the effect of miR-769-5p on the tumorigenicity of GC cells in vivo was investigated by nude mouse tumorigenicity assay. Moreover, the downstream target genes of miR-769-5p were predicted by bio-informatics. Finally, qRT-PCR and Western blotting were used to screen the downstream target genes.RESULTS: In the 62 cases of GC tissues, the expression of miR-769-5p was upregulated in 48 cases. MiR-769-5p was divided into high-expression group and low-expression group. Chi-square analysis showed that the high expression of miR-7695p was positively correlated with tumor-node-metastasis (TNM) stage (p=0.005), lymph node metastasis (p=0.010), and infiltration depth (p=0.011) in patients with GC. The results of qRT-PCR indicated that the expression of miR-769-5p was upregulated in GC cells. In vitro experiments (MIT, flow cytometry, EdU) results showed that after interfering in the expression of miR-769-5p, the proliferation ability of GC cells was decreased, and apoptosis was increased. Furthermore, the results of in vivo experiments manifested that the tumorigenic ability of GC cells declined after interference in the expression of miR-769-5p. Finally, the results of qRT-PCR and Western blotting revealed that the expression of RING1 and YY1-binding protein (RYBP) was regulated by miR-769-5p.CONCLUSIONS: The expression of miR-7695p is upregulated in GC and positively correlated with TNM stage in GC patients. By regulating the expression of RYBP, the proliferation of GC cells was promoted, and the apoptosis was inhibited.