Ai-lncRNA EGOT enhancing autophagy sensitizes paclitaxel cytotoxicity via upregulation of ITPR1 expression by RNA-RNA and RNA-protein interactions in human cancer
Ai-lncRNA EGOT enhancing autophagy sensitizes paclitaxel cytotoxicity via upregulation of ITPR1 expression by RNA-RNA and RNA-protein interactions in human cancer
复制标题
在人类癌症中，Ai-lncRNA EGOT 通过 RNA-RNA 和 RNA-蛋白质相互作用上调 ITPR1 表达来增强自噬，从而使紫杉醇细胞毒性敏感
DOI：
10.1186/s12943-019-1017-z
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发表时间：
2019-04-18
期刊：
影响因子：
37.3
通讯作者：
Pang, Da
中科院分区：
文献类型：
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作者：
Xu, Shouping;Wang, Peiyuan;Pang, Da
背景：反义内含子长链非编码RNA （Ai-lncRNA）的生物学功能尚不清楚。同时，临床上紫杉醇耐药的癌症患者治疗选择有限。然而，Ai-lncRNA在人类癌症紫杉醇敏感性中的潜在参与尚不清楚。方法：对33例乳腺标本进行全转录组测序，鉴定Ai-lncRNA EGOT。接下来，研究了EGOT在紫杉醇敏感性调控中的作用。此外，我们还详细研究了EGOT通过RNA-RNA和rna -蛋白相互作用上调ITPR1表达增强自噬增敏紫杉醇细胞毒性的机制。此外，通过免疫共沉淀法和染色质免疫共沉淀法研究了EGOT表达的上游转录调控。最后，通过本队列的临床乳腺标本、TCGA和ICGC来验证EGOT在增强紫杉醇敏感性中的作用。结果：EGOT通过上调ITPR1表达增强自噬体积累，从而使细胞对紫杉醇毒性敏感。从机制上讲，一方面，EGOT通过形成pre-ITPR1/EGOT dsRNA，诱导pre-ITPR1积累，增加cis中ITPR1蛋白的表达，从而上调ITPR1水平。另一方面，EGOT通过外显子1 EGOT第2段（324-645个核苷酸）的两个结合基序，招募hnRNPH1来增强trans中pre-ITPR1的选择性剪接。此外，EGOT受应激条件的转录调节。最后，通过评估癌症标本，EGOT表达增强了紫杉醇敏感性。结论：这些发现拓宽了对ai - lncrna生物学功能的全面认识。适当调节EGOT可能是提高紫杉醇敏感性的一种新的协同治疗策略。
Background:The biology function of antisense intronic long noncoding RNA (Ai-lncRNA) is still unknown. Meanwhile, cancer patients with paclitaxel resistance have limited therapeutic options in the clinic. However, the potential involvement of Ai-lncRNA in paclitaxel sensitivity remains unclear in human cancer.Methods:Whole transcriptome sequencing of 33 breast specimens was performed to identify Ai-lncRNA EGOT. Next, the role of EGOT in regulation of paclitaxel sensitivity was investigated. Moreover, the mechanism of EGOT enhancing autophagy sensitizes paclitaxel cytotoxicity via upregulation of ITPR1 expression by RNA-RNA and RNA-protein interactions was investigated in detail. Furthermore, upstream transcriptional regulation of EGOT expression was also investigated by co-immunoprecipitation and chromatin immunoprecipitation. Finally, clinical breast specimens in our cohort, TCGA and ICGC were applied to validate the role of EGOT in enhancing of paclitaxel sensitivity.Results:EGOT enhances autophagosome accumulation via the up-regulation of ITPR1 expression, thereby sensitizing cells to paclitaxel toxicity. Mechanistically, on one hand, EGOT upregulates ITPR1 levels via formation of a pre-ITPR1/EGOT dsRNA that induces pre-ITPR1 accumulation to increase ITPR1 protein expression in cis. On the other hand, EGOT recruits hnRNPH1 to enhance the alternative splicing of pre-ITPR1 in trans via two binding motifs in EGOT segment 2 (324-645 nucleotides) in exon 1. Moreover, EGOT is transcriptionally regulated by stress conditions. Finally, EGOT expression enhances paclitaxel sensitivity via assessment of cancer specimens.Conclusions:These findings broaden comprehensive understanding of the biology function of Ai-lncRNAs. Proper regulation of EGOT may be a novel synergistic strategy for enhancing paclitaxel sensitivity in cancer therapy.