Downregulation of miR-138 Sustains NF-κB Activation and Promotes Lipid Raft Formation in Esophageal Squamous Cell Carcinoma
Downregulation of miR-138 Sustains NF-κB Activation and Promotes Lipid Raft Formation in Esophageal Squamous Cell Carcinoma
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DOI：
10.1158/1078-0432.ccr-12-3169
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发表时间：
2013-03-01
影响因子：
11.5
通讯作者：
Li, Jun
中科院分区：
文献类型：
--
作者：
Gong, Hui;Song, Libing;Li, Jun
目的：核因子-kappaB信号的结构性激活在食管鳞癌的发生发展中起重要作用。本研究的目的是评价miR-138对核因子-kappaB的激活和ESCC进展的影响。实验设计：采用实时荧光定量聚合酶链式反应技术检测miR-138在ESCC细胞系、ESCC组织和205例ESSC标本中的表达。通过锚定非依赖性生长、鸡绒毛膜-尿囊膜、Transwell基质侵袭和Annexin V结合分析以及移植瘤模型来确定miR-138在ESCC进展中的作用。结果：miR-138在食管癌中表达下调，与肿瘤进展和患者生存呈负相关。在体外和体内，miR-138的下调增强了ESCC细胞的侵袭性，而miR-138的上调则降低了ESCC的侵袭性。沉默miR-138可促进K63连接的核因子-kappa B信号中间体TRAF2和RIP1的泛素化，并维持核因子-kappaB的激活。此外，miR-138的下调通过上调脂筏的多种成分，包括FLOT1、FLOT2和小窝蛋白-1来诱导脂筏的形成。重要的是，体外分析结果与一组人食管癌标本中miR-138的表达和NF-kB的过度激活之间存在显著的负相关。结论：我们的结果表明miR-138是一种抑癌的miRNA，miR-138的下调参与了结构性的NF-kappa B的激活和食管癌的进展。临床癌症资源；19(5)；1083-93。(C)2013年AACR。
Purpose: Constitutive activation of NF-kappa B signaling plays vital roles in esophageal squamous cell carcinoma (ESCC) progression. The aim of this study was to evaluate the effect of miR-138 on NF-kappa B activation and ESCC progression.Experimental Design: Expression of miR-138 in ESCC cell lines, ESCC tissues, and 205 archived ESSC specimens was determined using real-time PCR analysis. Anchorage-independent growth, chicken chorio-allantoic membrane, Transwell matrix invasion and Annexin V-binding assays, and a xenograft tumor model were used to determine the role of miR-138 in ESCC progression. The effect of miR-138 on NF-kappa B activation was investigated using IKK in vitro kinase, electrophoretic mobility shift, lipid raft isolation, and luciferase reporter assays.Results: miR-138 was downregulated and inversely correlated with tumor progression and patient survival in ESCCs. Downregulation of miR-138 enhanced, whereas upregulation of miR-138 reduced, the aggressive phenotype of ESCC cells both in vitro and in vivo. Silencing miR-138 promoted K63-linked polyubiquitination of the NF-kappa B signaling intermediaries TRAF2 and RIP1 and sustained NF-kappa B activation. Furthermore, downregulation of miR-138 induced lipid raft formation via upregulating multiple components of lipid rafts, including FLOT1, FLOT2, and caveolin-1. Importantly, the in vitro analysis was consistent with a significant inverse correlation between miR-138 expression and NF-kB hyperactivation in a cohort of human ESCC specimens.Conclusion: Our results show that miR-138 functions as a tumor-suppressive miRNA and that downregulation of miR-138 contributes to constitutive NF-kappa B activation and ESCC progression. Clin Cancer Res; 19(5); 1083-93. (C) 2013 AACR.