Long Non-coding RNA TPRG1-AS1 Interacts With CLTC in Liver Cancer Cells.

IF 1.6 4区医学 Q4 ONCOLOGY

Anticancer research Pub Date : 2024-11-01 DOI:10.21873/anticanres.17307

Sung Ung Moon, Masaud Shah, Trinh Thanh Thao, Hyun Goo Woo

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引用次数: 0

Abstract

Background/aim: Certain long non-coding RNAs (lncRNAs), identified as potential tumor suppressors, have shown potential in inhibiting tumor growth. Here, we investigated a novel mechanism involving the direct interaction between lncRNA TPRG1-AS1 and Clathrin Heavy Chain (CLTC) in the Epidermal Growth Factor (EGF) signaling pathway for its tumor-suppressive effects.

Materials and methods: Our research revealed a direct physical interaction between TPRG1-AS1 and CLTC through RNA pulldown and RNA immunoprecipitation (RIP)-qPCR, which subsequently influenced the EGF signaling pathway. We confirmed phenotype changes by cell viability, sphere formation, and invasion. We confirmed the mechanism underlying these phenotypic changes through immunoblotting and immunocytochemistry.

Results: Firstly, we confirmed a reduction in the phenotype associated with the overexpression of TPRG1-AS1 (TPRG1-AS1oe) interacting with CLTC in the presence of EGF signaling. Next, it was observed that TPRG1-AS1oe suppressed EGF downstream signaling, specifically MAPK8 and MAPK14, in relation to CLTC. Moreover, we verified that overexpressed TPRG1-AS1 binds to CLTC and suppresses EGF downstream signaling using a custom HEX probe.

Conclusion: Collectively our study uncovered a novel regulatory axis wherein TPRG1-AS1 interacts with CLTC, consequently attenuating EGF downstream signaling, particularly through the MAPK8 and MAPK14 pathways. These complex interactions ultimately lead to a reduction in the phenotype of liver cancer cells.

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长非编码 RNA TPRG1-AS1 与肝癌细胞中的 CLTC 相互作用

背景/目的：某些长非编码RNA（lncRNA）被认为是潜在的肿瘤抑制因子，在抑制肿瘤生长方面具有潜力。在此，我们研究了lncRNA TPRG1-AS1与表皮生长因子（EGF）信号通路中的Clathrin Heavy Chain（CLTC）直接相互作用的新机制，以了解其抑制肿瘤生长的作用：我们的研究通过RNA pulldown和RNA免疫沉淀（RIP）-qPCR发现了TPRG1-AS1和CLTC之间的直接物理相互作用，并进而影响了EGF信号通路。我们通过细胞活力、球体形成和侵袭证实了表型的变化。我们通过免疫印迹和免疫细胞化学证实了这些表型变化的机制：首先，我们证实了在存在 EGF 信号的情况下，TPRG1-AS1（TPRG1-AS1oe）与 CLTC 相互作用的表型减少。接着，我们观察到 TPRG1-AS1oe 抑制了与 CLTC 有关的 EGF 下游信号，特别是 MAPK8 和 MAPK14。此外，我们使用定制的HEX探针验证了过表达的TPRG1-AS1与CLTC结合并抑制EGF下游信号：总之，我们的研究发现了一个新的调控轴，即TPRG1-AS1与CLTC相互作用，从而抑制EGF下游信号传导，特别是通过MAPK8和MAPK14途径。这些复杂的相互作用最终导致肝癌细胞表型的减少。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Anticancer research 医学-肿瘤学

CiteScore

3.70

自引率

10.00%

发文量

566

审稿时长

2 months

期刊介绍： ANTICANCER RESEARCH is an independent international peer-reviewed journal devoted to the rapid publication of high quality original articles and reviews on all aspects of experimental and clinical oncology. Prompt evaluation of all submitted articles in confidence and rapid publication within 1-2 months of acceptance are guaranteed. ANTICANCER RESEARCH was established in 1981 and is published monthly (bimonthly until the end of 2008). Each annual volume contains twelve issues and index. Each issue may be divided into three parts (A: Reviews, B: Experimental studies, and C: Clinical and Epidemiological studies). Special issues, presenting the proceedings of meetings or groups of papers on topics of significant progress, will also be included in each volume. There is no limitation to the number of pages per issue.