Abnormal angiogenesis is crucial for gallbladder cancer (GBC) tumor growth and invasion, highlighting the importance of elucidating the mechanisms underlying this process. LncRNA (long non-coding RNA) is widely involved in the malignancy of GBC. However, conclusive evidence confirming the correlation between lncRNAs and angiogenesis in GBC is lacking. LncRNA sequencing was performed to identify the differentially expressed lncRNAs. RT-qPCR, western blot, FISH, and immunofluorescence were used to measure TRPM2-AS and NOTCH1 signaling pathway expression in vitro. Mouse xenograft and lung metastasis models were used to evaluate the biological function of TRPM2-AS during angiogenesis in vivo. EDU, transwell, and tube formation assays were used to detect the angiogenic ability of HUVECs. RIP, RAP, RNA pull-down, dual-luciferase reporter system, and mass spectrometry were used to confirm the interaction between TRPM2-AS, IGF2BP2, NUMB, and PABPC1. TRPM2-AS was upregulated in GBC tissues and was closely related to angiogenesis and poor prognosis in patients with GBC. The high expression level and stability of TRPM2-AS benefited from m6A modification, which is recognized by IGF2BP2. In terms of exerting pro-angiogenic effects, TRPM2-AS loaded with exosomes transported from GBC cells to HUVECs enhanced PABPC1-mediated NUMB expression inhibition, ultimately promoting the activation of the NOTCH1 signaling pathway. PABPC1 inhibited NUMB mRNA expression through interacting with AGO2 and promoted miR-31-5p and miR-146a-5p-mediated the degradation of NUMB mRNA. The NOTCH signaling pathway inhibitor DAPT inhibited GBC tumor angiogenesis, and TRPM2-AS knockdown enhanced this effect. TRPM2-AS is a novel and promising biomarker for GBC angiogenesis that promotes angiogenesis by facilitating the activation of the NOTCH1 signaling pathway. Targeting TRPM2-AS opens further opportunities for future GBC treatments.

中文翻译：

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外泌体长非编码RNA TRPM2-AS通过与PABPC1相互作用激活NOTCH1信号通路促进胆囊癌血管生成

异常血管生成对于胆囊癌（GBC）肿瘤的生长和侵袭至关重要，凸显了阐明这一过程背后机制的重要性。 LncRNA（长非编码RNA）广泛参与GBC的恶性肿瘤。然而，缺乏确凿的证据证实 lncRNA 与 GBC 中血管生成之间的相关性。进行lncRNA测序以鉴定差异表达的lncRNA。采用RT-qPCR、western blot、FISH和免疫荧光法检测体外TRPM2-AS和NOTCH1信号通路的表达。使用小鼠异种移植和肺转移模型来评估TRPM2-AS在体内血管生成过程中的生物学功能。 EDU、Transwell 和成管实验用于检测 HUVEC 的血管生成能力。使用 RIP、RAP、RNA pull-down、双荧光素酶报告系统和质谱来确认 TRPM2-AS、IGF2BP2、NUMB 和 PABPC1 之间的相互作用。 TRPM2-AS在GBC组织中表达上调，与GBC患者的血管生成和不良预后密切相关。 TRPM2-AS的高表达水平和稳定性得益于m6A修饰，该修饰被IGF2BP2识别。在发挥促血管生成作用方面，装载有从GBC细胞转运至HUVEC的外泌体的TRPM2-AS增强了PABPC1介导的NUMB表达抑制，最终促进NOTCH1信号通路的激活。 PABPC1 通过与 AGO2 相互作用抑制 NUMB mRNA 表达，并促进 miR-31-5p 和 miR-146a-5p 介导的 NUMB mRNA 降解。 NOTCH 信号通路抑制剂 DAPT 抑制 GBC 肿瘤血管生成，而 TRPM2-AS 敲低增强了这种作用。 TRPM2-AS 是一种新颖且有前途的 GBC 血管生成生物标志物，可通过促进 NOTCH1 信号通路的激活来促进血管生成。靶向 TRPM2-AS 为未来 GBC 治疗开辟了更多机会。