miR-194 targets RBX1 gene to modulate proliferation and migration of gastric cancer cells
miR-194 targets RBX1 gene to modulate proliferation and migration of gastric cancer cells
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DOI：
10.1007/s13277-014-2849-1
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发表时间：
2015-04-01
期刊：
影响因子：
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通讯作者：
Zhao, Guoqiang
中科院分区：
文献类型：
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作者：
Chen, Xiaonan;Wang, Yuanyuan;Zhao, Guoqiang
RING box蛋白1（RBX 1）是SCF E3泛素连接酶的重要组成部分，在胃癌发生发展中起重要作用。本研究采用qRT-PCR方法检测76对胃癌组织和非胃癌组织中miR-194和RBX 1的表达，并分析其临床病理特征。分别采用CCK 8、transwell法、伤口愈合法和流式细胞术检测miR-194对胃癌细胞增殖、侵袭、迁移、凋亡和细胞周期的影响。荧光素酶报告基因测定和蛋白质印迹法用于评估RBX 1是否是miR-194的直接靶标。采用Kaplan-Meier方法和log-rank检验评估miR-194或RBX 1表达与患者生存期之间的相关性。然后，我们发现miR-194在GC组织中显著下调，RBX 1上调;两者均与肿瘤大小，位置，侵袭和肿瘤淋巴结转移显著相关。miR-194过表达显著限制了细胞增殖、侵袭和迁移。miR-194下调RBX 1蛋白表达，荧光素酶测定显示RBX 1 3 'UTR中的结合位点是miR-194介导的RBX 1抑制所必需的，表明RBX 1是miR-194的直接靶点。无3 'UTR的RBX 1的转染恢复了miR-194抑制迁移功能。miR-194过表达或RBX 1低表达与胃癌患者生存期延长相关。总之，上调miR-194可能通过靶向RBX 1抑制GC细胞的增殖、迁移和侵袭。miR-194和RBX 1的异常表达与胃癌患者的生存时间相关。
RING box protein1 (RBX1), an essential component of SCF E3 ubiquitin ligases, plays an important role in gastric cancer. In the study, miR-194 and RBX1 expression was evaluated in 76 pairs of gastric tumor and non-tumor tissue samples by qRT-PCR, and clinicopathological characteristics were analyzed. CCK8, transwell assay, wound healing assay, and flow cytometry assay were performed to evaluate the effect of miR-194 on gastric cancer (GC) cellular proliferation, invasion, migration, apoptosis, and cell cycle, respectively. Luciferase reporter assays and Western blotting were used to evaluate whether RBX1 is a direct target of miR-194. The Kaplan-Meier method and log-rank test were used to evaluate the correlation between miR-194 or RBX1 expression and patient survival. Then, we found that miR-194 was significantly downregulated and RBX1 upregulated in GC tissues; both of which showed significant association with tumor size, location, invasion, and tumor node metastasis. Cell proliferation, invasion, and migration were significantly restricted with miR-194 overexpression. miR-194 downregulated RBX1 protein expression, and luciferase assays showed that binding sites in the RBX1 3'UTR were required for miR-194-mediated repression of RBX1, indicating that RBX1 was a direct target of miR-194. Transfection of RBX1 without the 3'UTR restored the miR-194-inhibiting migration function. miR-194 overexpression or RBX1 lowexpression was associated with prolonged survival of GC patients. In conclusion, upregulation of miR-194 can inhibit proliferation, migration, and invasion of GC cells, possibly by targeting RBX1. Aberrant expression of miR-194 and RBX1 is correlated to GC patient survival time.