miR-769-5p 通过下调胃癌中 RYBP 的表达发挥癌基因作用。
MiR-769-5p functions as an oncogene by down-regulating RYBP expression in gastric cancer.
机构信息
Department of General Surgery, Yantaishan Hospital, Yantai, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Jun;24(12):6699-6706. doi: 10.26355/eurrev_202006_21657.
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 bioinformatics. 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-769-5p 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 (MTT, 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-769-5p 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.
目的
本研究旨在检测微小 RNA（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 GC 中的临床意义、生物学功能和作用机制。
方法
采用实时荧光定量聚合酶链反应（qRT-PCR）检测 62 例 GC 组织和癌旁组织中 miR-769-5p 的相对表达水平。采用卡方检验分析 miR-769-5p 表达与 GC 患者临床病理特征的相关性。此外，采用 qRT-PCR 检测 GC 细胞中 miR-769-5p 的相对表达水平和 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。接下来，通过裸鼠肿瘤生成实验研究 miR-769-5p 对 GC 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 Western blot 筛选下游靶基因。
结果
在 62 例 GC 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。卡方分析表明，miR-769-5p 的高表达与 GC 患者的肿瘤-淋巴结-转移（TNM）分期（p=0.005）、淋巴结转移（p=0.010）和浸润深度（p=0.011）呈正相关。qRT-PCR 结果表明，GC 细胞中 miR-769-5p 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，GC 细胞的增殖能力降低，凋亡增加。此外，体内实验结果表明，干扰 miR-769-5p 表达后，GC 细胞的致瘤能力下降。最后，qRT-PCR 和 Western blot 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 GC 中表达上调，与 GC 患者的 TNM 分期呈正相关。通过调节 RYBP 的表达，促进了 GC 细胞的增殖，抑制了细胞凋亡。
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