Long non-coding RNA NORAD/miR-224-3p/MTDH axis contributes to CDDP resistance of esophageal squamous cell carcinoma by promoting nuclear accumulation of β-catenin.
Long non-coding RNA NORAD/miR-224-3p/MTDH axis contributes to CDDP resistance of esophageal squamous cell carcinoma by promoting nuclear accumulation of β-catenin.
复制标题
长非编码 RNA NORAD/miR-224-3p/MTDH 轴通过促进 β-catenin 的核积累有助于食管鳞状细胞癌的 CDDP 抵抗
DOI：
10.1186/s12943-021-01455-y
复制
发表时间：
2021-12-10
期刊：
影响因子：
37.3
通讯作者：
Liu L
中科院分区：
文献类型：
--
作者：
Jia Y;Tian C;Wang H;Yu F;Lv W;Duan Y;Cheng Z;Wang X;Wang Y;Liu T;Wang J;Liu L
背景以顺铂（CDDP）为基础的化疗方案是食管鳞癌（ESCC）的主要治疗方案。失调的长非编码RNA（lncRNA）有助于CDDP抗性，这导致ESCC患者的治疗失败。然而，大多数的lncRNA参与CDDP耐药ESCC仍有待阐明。MethodsThe公共基因表达Omnibus（GEO）数据集GSE 45670进行了分析，以揭示潜在的lncRNA参与CDDP耐药ESCC。通过qRT-PCR在ESCC标本中检测候选上调的lncRNA以鉴定关键的lncRNA。选择DNA损伤激活的非编码RNA（NORAD）进行进一步研究。Kaplan-Meier分析和考克斯比例回归模型分析NORAD预测ESCC患者预后的潜力。NORAD在CDDP抗性中的作用通过进行体外功能获得和丧失实验来确定。荧光原位杂交（FISH），以确定NORAD在食管鳞癌细胞的亚细胞定位。一个公共的GEO数据集和生物信息学算法被用来预测可能被NORAD潜在地吸收的microRNA（miRNAs）。进行qRT-PCR以验证候选miRNA的表达。进行荧光素酶报告基因和Argonaute-2（Ago 2）-RNA免疫沉淀（RIP）测定以评估NORAD和候选miRNA之间的相互作用。进行miRNA拯救实验以验证NORAD调节轴及其对ESCC细胞中CDDP抗性的影响。进行蛋白质印迹以确认NORAD的精确下游信号传导途径。NORAD在CDDP耐药的食管鳞癌组织和细胞中的表达高于CDDP敏感的组织和细胞。NORAD表达与接受CDDP化疗的ESCC患者的术后预后呈负相关。NORAD敲除部分地阻止了ESCC细胞的CDDP抗性。FISH显示NORAD定位于食管鳞癌细胞的胞浆中。此外，生物信息学算法分析和qRT-PCR的重叠结果表明，NORAD可以在ESCC细胞中海绵miR-224- 3 p。Ago 2-RIP证实NORAD和miR-224- 3 p占据相同的Ago 2形成RNA诱导沉默复合物（RISC），并随后调节ESCC细胞中metadherin（MTDH）的表达。NORAD/miR-224- 3 p/MTDH轴通过促进β-catenin在细胞核内的积累，促进食管鳞癌细胞对顺铂的耐药和进展。我们的研究结果强调了NORAD作为接受CDDP化疗的ESCC患者治疗靶点的潜力。
BackgroundCis-diamminedichloro-platinum (CDDP)-based chemotherapy regimens are the most predominant treatment strategies for patients with esophageal squamous cell carcinoma (ESCC). Dysregulated long non-coding RNAs (lncRNAs) contribute to CDDP resistance, which results in treatment failure in ESCC patients. However, the majority of lncRNAs involved in CDDP resistance in ESCC remain to be elucidated.MethodsThe public Gene Expression Omnibus (GEO) dataset GSE45670 was analysed to reveal potential lncRNAs involved in CDDP resistance of ESCC. Candidate upregulated lncRNAs were detected in ESCC specimens by qRT-PCR to identify crucial lncRNAs. Non-coding RNA activated by DNA damage (NORAD) was selected for further study. Kaplan-Meier analysis and a COX proportional regression model were performed to analyse the potential of NORAD for predicting prognosis of ESCC patients. The role of NORAD in CDDP resistance were determined by conducting gain and loss-of-function experiments in vitro. Fluorescence in situ hybridization (FISH) was performed to determine the subcellular location of NORAD in ESCC cells. A public GEO dataset and bioinformatic algorithms were used to predict the microRNAs (miRNAs) that might be latently sponged by NORAD. qRT-PCR was conducted to verify the expression of candidate miRNAs. Luciferase reporter and Argonaute-2 (Ago2)-RNA immunoprecipitation (RIP) assays were conducted to evaluate the interaction between NORAD and candidate miRNAs. A miRNA rescue experiment was performed to authenticate the NORAD regulatory axis and its effects on CDDP resistance in ESCC cells. Western blotting was conducted to confirm the precise downstream signalling pathway of NORAD. A xenograft mouse model was established to reveal the effect of NORAD on CDDP resistance in vivo.ResultsThe expression of NORAD was higher in CDDP-resistant ESCC tissues and cells than in CDDP-sensitive tissues and cells. NORAD expression was negatively correlated with the postoperative prognosis of ESCC patients who underwent CDDP-based chemotherapy. NORAD knockdown partially arrested CDDP resistance of ESCC cells. FISH showed that NORAD was located in the cytoplasm in ESCC cells. Furthermore, overlapping results from bioinformatic algorithms analyses and qRT-PCR showed that NORAD could sponge miR-224-3p in ESCC cells. Ago2-RIP demonstrated that NORAD and miR-224-3p occupied the same Ago2 to form an RNA-induced silencing complex (RISC) and subsequently regulated the expression of metadherin (MTDH) in ESCC cells. The NORAD/miR-224-3p/MTDH axis promoted CDDP resistance and progression in ESCC cells by promoting nuclear accumulation of β-catenin in vitro and in vivo.ConclusionsNORAD upregulates MTDH to promote CDDP resistance and progression in ESCC by sponging miR-224-3p. Our results highlight the potential of NORAD as a therapeutic target in ESCC patients receiving CDDP-based chemotherapy.
登录
查看更多内容
影响因子：
3.5
作者：
Khan M;Sarkar D
通讯作者：
Sarkar D
DOI：
10.1016/s0140-6736(17)33326-3
发表时间：
2018-03-17
期刊：
Lancet (London, England)
影响因子：
--
作者：
Allemani C;Matsuda T;Di Carlo V;Harewood R;Matz M;Nikšić M;Bonaventure A;Valkov M;Johnson CJ;Estève J;Ogunbiyi OJ;Azevedo E Silva G;Chen WQ;Eser S;Engholm G;Stiller CA;Monnereau A;Woods RR;Visser O;Lim GH;Aitken J;Weir HK;Coleman MP;CONCORD Working Group
通讯作者：
CONCORD Working Group
星形胶质细胞升高基因-1（AEG-1）在癌症中的多功能作用：关注耐药性。
影响因子：
5.2
作者：
Manna D;Sarkar D
通讯作者：
Sarkar D
LncRNA-TUSC7/miR-224通过竞争性调节DESC1影响食管鳞癌化疗耐药
DOI：
10.1186/s13046-018-0724-4
发表时间：
2018-03-12
期刊：
Journal of experimental & clinical cancer research : CR
影响因子：
--
作者：
Chang ZW;Jia YX;Zhang WJ;Song LJ;Gao M;Li MJ;Zhao RH;Li J;Zhong YL;Sun QZ;Qin YR
通讯作者：
Qin YR
影响因子：
6.4
作者：
Jia, Yunlong;Wang, Hongyan;Liu, Lihua
通讯作者：
Liu, Lihua