LncRNA MIR4435-2HG drives cancer progression by modulating cell cycle regulators and mTOR signaling in stroma-enriched subtypes of urothelial carcinoma of the bladder.
LncRNA MIR4435-2HG drives cancer progression by modulating cell cycle regulators and mTOR signaling in stroma-enriched subtypes of urothelial carcinoma of the bladder.
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DOI：
10.1007/s13402-023-00826-5
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发表时间：
2023-10
影响因子：
6.6
通讯作者：
Qin, Haide
中科院分区：
文献类型：
--
作者：
Pei, Lu;Yan, Dong;He, Qingqing;Kong, Jianqiu;Yang, Meihua;Ruan, Honglian;Lin, Qiongqiong;Huang, Lifang;Huang, Jian;Lin, Tianxin;Qin, Haide
膀胱尿路上皮癌（UCB）治疗后复发和转移的风险较高。因此，迫切需要鉴定有效的预后标志物和新的治疗靶点。一些长链非编码RNA（lncRNA）已被报道与UCB进展相关。在本研究中，我们发现亚型特异性lncRNA MIR 4435 -2宿主基因（MIR 4435 - 2 HG）在UCB中发挥新的致癌作用。分析TCGA/BLCA的RNA-Seq数据。采用qRT-PCR检测16对膀胱癌组织及癌旁正常组织中MIR 4435 - 2 HG的表达。MIR 4435 - 2 HG的临床相关性通过对116个UCB患者样本的内部队列进行原位杂交来验证。进行RNA下拉，然后进行质谱分析以鉴定MIR 4435 - 2 HG结合蛋白。为了鉴定参与MIR 4435 - 2 HG活性的信号传导途径，使用其中MIR 4435 - 2 HG表达被敲低或外源性过表达的UCB细胞进行全面的体外和体内研究和RNA-Seq测定。此外，我们进行了RNA免疫沉淀和Western印迹分析，以验证所鉴定的MIR 4435 - 2 HG结合蛋白，并确定MIR 4435 - 2 HG促进UCB进展的分子机制。我们发现MIR 4435 - 2 HG在UCB的基质富集亚型中显著上调。MIR 4435 - 2 HG的高表达与高组织学分级、晚期T分期、较大肿瘤、淋巴结转移和不良预后呈正相关。体外实验显示MIR 4435 - 2 HG表达沉默抑制细胞增殖并诱导细胞凋亡。MIR 4434 - 2 HG的抑制延迟了异种移植肿瘤的生长，而MIR 4435 - 2 HG过表达逆转了MIR 4435 - 2 HG沉默诱导的UCB肿瘤表型获得的抑制。从机制上讲，我们发现MIR 4435 - 2 HG正调控多种细胞周期调节因子的表达，包括BRCA 2和CCND 1。敲低MIR 4435 - 2 HG增加了肿瘤细胞对VEGFR抑制剂西地尼布的敏感性。此外，我们发现MIR 4435 - 2 HG通过调节mTOR、70 S6 K和4 EBP 1的磷酸化来调节mTOR信号传导和上皮-间质转化（EMT）信号传导途径。最后，我们证实MIR 4435 - 2 HG通过调节EMT途径增强肿瘤转移。我们的数据表明，上调的MIR 4435 - 2 HG表达水平与UCB患者的不良预后显著相关。MIR 4435 - 2 HG促进膀胱癌进展，介导细胞周期（去）调节并调节mTOR信号传导。MIR 4435 - 2 HG是UCB中的致癌lncRNA，可用作诊断和治疗靶标。在线版本包含补充材料，可通过10.1007/s13402-023-00826-5获得。
The risk for recurrence and metastasis after treatment for urothelial carcinoma of the bladder (UCB) is high. Therefore, identifying efficient prognostic markers and novel therapeutic targets is urgently needed. Several long noncoding RNAs (lncRNAs) have been reported to be correlated with UCB progression. In this study, we found that the subtype-specific lncRNA MIR4435-2 host gene (MIR4435-2HG) plays a novel oncogenic role in UCB. RNA-Seq data of TCGA/BLCA were analyzed. The expression of MIR4435-2HG was measured by qRT-PCR in 16 pairs of bladder cancer tissues and adjacent normal tissues. The clinical relecance of MIR4435-2HG was validated via in situ hybridization performed on an in-house cohort of 116 UCB patient samples. RNA pull-down followed by mass spectrometry was performed to identify MIR4435-2HG-binding proteins. To identify signaling pathways involved in MIR4435-2HG activity, comprehensive in vitro and in vivo studies and RNA-Seq assays were performed using UCB cells in which MIR4435-2HG expression was knocked down or exogenously overexpressed. In addition, we performed RNA immunoprecipitation and Western blot analyses to validate the identified MIR4435-2HG-binding proteins and to determine the molecular mechanisms by which MIR4435-2HG promotes UCB progression. We found that MIR4435-2HG was significantly upregulated in the stromal-enriched subtype of UCB. Increased MIR4435-2HG expression was positively correlated with a high histological grade, advanced T stages, larger tumors, lymph node metastasis and a poor prognosis. In vitro experiments revealed that MIR4435-2HG expression silencing suppressed cell proliferation and induced apoptosis. Inhibition of MIR4434-2HG delayed xenograft tumor growth, while MIR4435-2HG overexpression reversed the MIR4435-2HG silencing-induced inhibition of UCB tumor phenotype acquisition. Mechanistically, we found that MIR4435-2HG positively regulated the expression of a variety of cell cycle regulators, including BRCA2 and CCND1. Knocking down MIR4435-2HG increased the sensitivity of tumor cells to the VEGFR inhibitor cediranib. Furthermore, we found that MIR4435-2HG regulated mTOR signaling and epithelial-mesenchymal transition (EMT) signaling pathways by modulating the phosphorylation of mTOR, 70S6K and 4EBP1. Finally, we confirmed that MIR4435-2HG enhances tumor metastasis through regulation of the EMT pathway. Our data indicate that upregulated MIR4435-2HG expression levels are significantly correlated with a poor prognosis of UCB patients. MIR4435-2HG promotes bladder cancer progression, mediates cell cycle (de)regulation and modulates mTOR signaling. MIR4435-2HG is an oncogenic lncRNA in UCB that may serve as a diagnostic and therapeutic target. The online version contains supplementary material available at 10.1007/s13402-023-00826-5.
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影响因子：
11.1
作者：
Bonasio R;Shiekhattar R
通讯作者：
Shiekhattar R
影响因子：
3.5
作者：
通讯作者：
--
影响因子：
64.5
作者：
Lee DF;Su J;Kim HS;Chang B;Papatsenko D;Zhao R;Yuan Y;Gingold J;Xia W;Darr H;Mirzayans R;Hung MC;Schaniel C;Lemischka IR
通讯作者：
Lemischka IR
DOI：
10.1093/jnci/djw302
发表时间：
2017-07-01
期刊：
Journal of the National Cancer Institute
影响因子：
--
作者：
Kuchenbaecker KB;McGuffog L;Barrowdale D;Lee A;Soucy P;Dennis J;Domchek SM;Robson M;Spurdle AB;Ramus SJ;Mavaddat N;Terry MB;Neuhausen SL;Schmutzler RK;Simard J;Pharoah PDP;Offit K;Couch FJ;Chenevix-Trench G;Easton DF;Antoniou AC
通讯作者：
Antoniou AC
影响因子：
12.4
作者：
Liu, Hai-Ting;Zou, Yong-Xin;Gao, Peng
通讯作者：
Gao, Peng