Identification of miR-29c and its Target FBXO31 as a Key Regulatory Mechanism in Esophageal Cancer Chemoresistance: Functional Validation and Clinical Significance
Identification of miR-29c and its Target FBXO31 as a Key Regulatory Mechanism in Esophageal Cancer Chemoresistance: Functional Validation and Clinical Significance
复制标题
miR-29c及其靶标FBXO31鉴定为食管癌化疗耐药关键调控机制——功能验证及临床意义
DOI：
10.7150/thno.30372
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发表时间：
2019-01-01
期刊：
影响因子：
12.4
通讯作者：
Cheung, Annie L. M.
中科院分区：
文献类型：
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作者：
Li, Bin;Hong, Pan;Cheung, Annie L. M.
原理：癌症中微RNA（miRNA）表达失调可能导致化疗耐药性。本研究旨在鉴定与食管鳞状细胞癌（ESCC）氟尿嘧啶（5-FU）化疗耐药相关的miRNA。探讨了miR-29 c作为ESCC新的诊断、预后和治疗预测标志物的潜力，以及其克服5-FU化疗耐药的机制和治疗意义。研究方法：利用Taqman miRNA微阵列分析获得性5-FU耐药ESCC细胞模型的miRNA谱，以鉴定与5-FU耐药相关的新miRNA。使用定量实时PCR来确定患者组织和血清样品中的miR-29 c表达。进行生物信息学、功能获得和丧失实验以及荧光素酶报告基因测定以验证F-box only蛋白31（FBXO 31）作为miR-29 c的直接靶点，并鉴定控制miR-29 c表达的潜在转录因子结合事件。在肿瘤异种移植模型中评估了基于miR-29 c阿糖胞苷的全身性疗法克服5-FU化疗耐药性的潜力。结果如下：miR-29 c在STAT 5A的调控下，在ESCC患者的肿瘤和血清样品中频繁下调，并且表达水平与总生存期相关。功能研究表明，miR-29 c可通过直接与FBXO 31的3 'UTR相互作用，抑制FBXO 31的表达并激活下游p38 MAPK，从而在体外和体内克服5-FU化疗耐药性。全身给予miR-29 c显著改善了体内5-FU化疗耐药ESCC异种移植物的反应。结论：MiR-29 c通过与FBXO 31相互作用调节化疗耐药性，是一种有前景的非侵袭性生物标志物和治疗ESCC的靶点。
Rationale: Dysregulated microRNA (miRNA) expressions in cancer can contribute to chemoresistance. This study aims to identify miRNAs that are associated with fluorouracil (5-FU) chemoresistance in esophageal squamous cell carcinoma (ESCC). The potential of miR-29c as a novel diagnostic, prognostic and treatment-predictive marker in ESCC, and its mechanisms and therapeutic implication in overcoming 5-FU chemoresistance were explored. Methods: The miRNA profiles of an ESCC cell model with acquired chemoresistance to 5-FU were analyzed using a Taqman miRNA microarray to identify novel miRNAs associated with 5-FU chemoresistance. Quantitative real-time PCR was used to determine miR-29c expression in tissue and serum samples of patients. Bioinformatics, gain- and loss-of-function experiments, and luciferase reporter assay were performed to validate F-box only protein 31 (FBXO31) as a direct target of miR-29c, and to identify potential transcription factor binding events that control miR-29c expression. The potential of systemic miR-29c oligonucleotide-based therapy in overcoming 5-FU chemoresistance was evaluated in tumor xenograft model. Results: MiR-29c, under the regulatory control of STAT5A, was frequently downregulated in tumor and serum samples of patients with ESCC, and the expression level was correlated with overall survival. Functional studies showed that miR-29c could override 5-FU chemoresistance in vitro and in vivo by directly interacting with the 3'UTR of FBXO31, leading to repression of FBXO31 expression and downstream activation of p38 MAPK. Systemically administered miR-29c dramatically improved response of 5-FU chemoresistant ESCC xenografts in vivo. Conclusions: MiR-29c modulates chemoresistance by interacting with FBXO31, and is a promising non-invasive biomarker and therapeutic target in ESCC.