| 摘要: |
| 目的:越来越多的证据表明,长链非编码RNA(lncRNA)参与了肝癌的发生发展。然而,lncRNA核仁RNA宿主基因3(SNHG3)在肝细胞癌中的致癌机制研究知之甚少。本研究旨在探究SNHG3是否调控miR-186-5p/E盒结合锌指蛋白1(ZEB1)促进肝癌细胞的迁移、侵袭和上皮-间充质转化。方法:在HepG2细胞中转染siRNA干扰片段敲除SNHG3,或转染pcDNA3.1+/SNHG3使SNHG3过表达,通过实时荧光定量PCR(qRT-PCR)检测是否转染成功。克隆形成实验检测HepG2细胞增殖；细胞划痕实验检测HepG2细胞迁移率；Transwell小室实验检测HepG2细胞侵袭数量。双荧光素酶活性基因检测验证SNHG3与miR-186-5p或miR-186-5p与ZEB1的靶向关系。RT-PCR和Western blot检测SNHG3、miR-186-5p、ZEB1、E-cadherin、N-cadherin、MMP2、Vimentin、MMP9和Snail的mRNA和蛋白相对表达量。结果:结果显示,肝癌组织中SNHG3的表达量显著高于正常组织,且SNHG3表达量高的患者生存率显著性低于SNHG3表达量低的患者(p=0.00033)。SNHG3过表达可诱导ZEB1过表达从而促进HepG2细胞增殖、侵袭、迁移和上皮-间充质转化,SNHG3敲除可下调ZEB1的表达从而抑制HepG2细胞侵袭、迁移和上皮-间充质转化。结论:SNHG3通过调控miR-186-5p/ZEB1促进肝癌细胞的迁移、侵袭和上皮-间充质转化,表明SNHG3是肝癌潜在的治疗靶点。 |
| 关键词: 肝癌 长链非编码RNA ZEB1 侵袭 迁移 上皮-间充质转化 |
| DOI： |
| 分类号:R735.7 |
| 基金项目: |
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| SNHG3 promotes migration, invasion, and epithelial-mesenchymal transition of hepatocellular carcinoma cells through the miR-186-5p/ZEB1 axisCHEN Xiaolan, SU Yayong, LIU Shangping, WANG Lihui, XU Chengrun#(The 909th Hospital of the People"s Liberation Army, Zhangzhou, 363100; #Corresponding Author:xcrun18@sina.com) |
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陈小兰
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联勤保障部队第九〇九医院（厦门大学附属东南医院）
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| Abstract: |
| Objective Increasing evidence has shown that long non-coding RNA(LncRNA) is involved in the occurrence and development of hepatocellular carcinoma(HCC). However, little is known about the oncogenic mechanism of lncRNA small nucleolar RNA host gene 3 (SNHG3) in HCC. The aim of this study is to investigate whether SNHG3 promotes the migration, invasion and epithelial-mesenchymal transition of HCC by regulating miR-186-5p/zinc finger E-box binding homeobox 1(ZEB1). Methods HepG2 cells were transfected with small interfering RNA (siRNA) to knock down SNHG3 or pcDNA3.1+/SNHG3 to overexpress SNHG3, and real-time fluorescence quantitative PCR (qRT-PCR) was used to detect whether the transfection was successful. HepG2 cell proliferation was detected by colony formation assay. Scratch wound healing assay was used to detect the migration rate of HepG2 cells. Transwell assay was used to detect the number of HepG2 cell invasion. Dual-luciferase reporter assay was used to verify the targeting relationship between SNHG3 and miR-186-5p or miR-186-5p and ZEB1. qRT-PCR and Western blot were used to detect the mRNA and protein expressions of SNHG3, miR-186-5p, ZEB1, E-cadherin, N-cadherin, MMP2, Vimentin, MMP9 and Snail. Results The results showed that the expression of SNHG3 in HCC tissues was significantly higher than that in normal tissues, and the survival rate of patients with high SNHG3 expression was significantly lower than that of patients with low SNHG3 expression (p=0.00033). Overexpression of SNHG3 induced overexpression of ZEB1 to promote the proliferation, invasion, migration and epithelial-mesenchymal transition of HepG2 cells, while SNHG3 knockout down-regulated the expression of ZEB1 to inhibit the invasion, migration and epithelial-mesenchymal transition of HepG2 cells. Conclusion SNHG3 promotes the migration, invasion and epithelial-mesenchymal transition of hepatocellular carcinoma cells by regulating miR-186-5p/ZEB1, indicating that SNHG3 is a potential treatment target for HCC. |
| Key words: HCC LncRNA ZEB1 Invasion Migration Epithelial-mesenchymal transition |
