Long Non-Coding RNA MDFIC-7 Promotes Chordoma Progression Through Modulating the miR-525-5p/ARF6 Axis.
Long Non-Coding RNA MDFIC-7 Promotes Chordoma Progression Through Modulating the miR-525-5p/ARF6 Axis.
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DOI：
10.3389/fonc.2021.743718
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发表时间：
2021
影响因子：
4.7
通讯作者：
Chen K
中科院分区：
文献类型：
--
作者：
Zhang K;Liu Z;Wang Z;Zhou Z;Shao X;Hua X;Mao H;Yang H;Ren K;Chen K
软骨瘤是一种极为罕见的恶性肿瘤，因其局部侵袭性强、复发率高而难以治愈。长链非编码RNA（lncRNA）已被证明在各种癌症中发挥多种作用。本研究的目的是研究lncRNA MDFIC-7在脉络膜中的调节功能，并阐明其潜在的机制。采用实时定量聚合酶链反应（RT-PCR）检测15例脉络膜癌组织、癌旁组织及脉络膜细胞系中lncRNA MDFIC-7的表达。通过RNA干扰和慢病毒转导进行基因沉默和过表达实验。通过细胞计数kit-8法、集落形成法和异种移植瘤实验评价lncRNA MDFIC-7对脉络膜细胞增殖的影响。采用RNA免疫沉淀和双荧光素酶报告基因分析方法，研究lncRNA MDFIC-7与miRNA-525 - 5 p的结合以及miR-525- 5 p与ADP-核糖基化因子6（ARF 6）mRNA 3′非翻译区的相互作用。用XF 96型细胞外液流量分析仪测定细胞糖酵解能力和线粒体功能。IncRNA MDFIC-7在脉络膜肿瘤组织中的表达高于癌旁组织。在裸鼠模型中，下调lncRNA MDFIC-7减少了脉络膜细胞中的集落形成和细胞增殖，并减少了异种移植肿瘤的生长。此外，lncRNA MDFIC-7敲低减弱了脉络膜细胞和异种移植肿瘤中的瓦尔堡效应。LncRNA MDFIC-7敲低升高了miR-525- 5 p水平并降低了ARF 6表达。ARF 6的过表达逆转了lncRNA MDFIC-7敲低对细胞增殖的抑制作用以及脉络膜细胞和异种移植肿瘤中的瓦尔堡效应。在机制上，lncRNA MDFIC-7作为miR-525- 5 p的分子海绵，负调控miR-525- 5 p的表达，并促进miR-525- 5 p靶点ARF 6的基因表达。我们的研究结果表明，lncRNA MDFIC-7作为一个分子海绵竞争性结合miR-525- 5 p和促进ARF 6的表达。lncRNA MDFIC-7/miR-525- 5 p/ARF 6轴调节脉络膜进展和脉络膜中的瓦尔堡效应，表明lncRNA MDFIC-7和miR-525- 5 p可能是治疗脉络膜的有希望的治疗靶点。
Chordoma, an extremely rare malignant tumor, remains difficult to be cured because of its strong local invasiveness and high recurrence rate. Long non-coding RNAs (lncRNAs) have been demonstrated to play multiple roles in various cancers. The purpose of this study was to investigate the modulatory function of lncRNA MDFIC-7 in chordoma and to elucidate its underlying mechanisms. Quantitative real-time polymerase chain reaction was performed to detect the expression of lncRNA MDFIC-7 in tumor tissues and adjacent nontumorous tissues collected from 15 chordoma patients, as well as in chordoma cell lines. Gene silencing and overexpression experiments were carried out by RNA interference and lentiviral transduction. The effect of lncRNA MDFIC-7 on the proliferation of chordoma cells was evaluated by cell counting kit-8 assay, colony formation assay and xenograft tumor experiments. RNA immunoprecipitation and dual luciferase reporter assays were conducted to evaluate the binding between lncRNA MDFIC-7 and miRNA-525-5p and the interaction between miR-525-5p and the 3′ untranslated region of ADP-ribosylation factor 6 (ARF6) mRNA. The glycolytic capacity and mitochondrial function of chordoma cells were measured by the Seahorse Bioscience XF96 Extracellular Flux Analyzer. The expression of lncRNA MDFIC-7 was higher in chordoma tumor tissues than in adjacent non-tumor tissues. Downregulation of lncRNA MDFIC-7 reduced colony formation and cell proliferation in chordoma cells and decreased xenograft tumor growth in a nude mouse model. Moreover, lncRNA MDFIC-7 knockdown attenuated the Warburg effect in chordoma cells and xenograft tumors. LncRNA MDFIC-7 knockdown elevated miR-525-5p levels and decreased ARF6 expressions. Overexpression of ARF6 reversed the inhibitory effect of lncRNA MDFIC-7 knockdown on cell proliferation and the Warburg effect in chordoma cells and xenograft tumors. Mechanistically, lncRNA MDFIC-7, as a molecular sponge of miR-525-5p, negatively regulated miR-525-5p expression and promoted the gene expression of ARF6, a miR-525-5p target. Our findings demonstrate that lncRNA MDFIC-7 acts as a molecular sponge to competitively bind to miR-525-5p and promote expression of ARF6. The lncRNA MDFIC-7/miR-525-5p/ARF6 axis regulates chordoma progression and the Warburg effect in chordoma, suggesting that lncRNA MDFIC-7 and miR-525-5p could be promising therapeutic targets for the treatment of chordoma.
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影响因子：
2.2
作者：
Chen, K. W.;Yang, H. L.;Chen, X. Q.
通讯作者：
Chen, X. Q.
影响因子：
16.6
作者：
Onodera Y;Nam JM;Horikawa M;Shirato H;Sabe H
通讯作者：
Sabe H
DOI：
10.1038/nrm3117
发表时间：
2011-06
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
--
影响因子：
4.9
作者：
Kreshak, Jennifer;Larousserie, Frederique;Vanel, Daniel
通讯作者：
Vanel, Daniel
影响因子：
3.5
作者：
Oka, Soichi;Uramoto, Hidetaka;Tanaka, Funnihiro
通讯作者：
Tanaka, Funnihiro