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敲低 ANLN 通过细胞焦亡抑制肺腺癌的进展。

Knockdown of ANLN inhibits the progression of lung adenocarcinoma via pyroptosis activation.

机构信息

Department of Medical Oncology, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, Hainan 570311, P.R. China.

Department of Psychology, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, Hainan 570311, P.R. China.

出版信息

Mol Med Rep. 2023 Sep;28(3). doi: 10.3892/mmr.2023.13064. Epub 2023 Aug 4.

Abstract

Significant advancements have been achieved in the area of molecular targeted therapy for lung adenocarcinoma (LUAD). However, the complex molecular patterns and high heterogeneity of LUAD confine the efficacy of these therapies to a specific subset of patients; therefore, it is necessary to explore novel targets for LUAD treatment. The expression levels of anillin (ANLN) in LUAD were analyzed using the Gene Expression Profiling Interactive Analysis database. Furthermore, the association between ANLN gene expression and patient survival outcomes was evaluated using the Kaplan‑Meier Plotter. Subsequently, small interfering RNA (siRNA) transfection was performed to knock down ANLN in A549 and H1299 cell lines, after which, TUNEL, colony formation and Transwell assays were conducted to assess cell death, colony formation and migration, respectively. Additionally, western blot analysis was performed to analyze the expression levels of caspase‑1, interleukin (IL)‑18 (IL‑18), IL‑1β, NLR family pyrin domain‑containing 3 (NLRP3), apoptosis‑associated speck‑like protein containing a CARD domain (ASC) and cleaved gasdermin D (GSDMD) following ANLN knockdown. The results revealed that ANLN mRNA expression was significantly increased in LUAD tissues compared with adjacent normal samples. Furthermore, the expression levels of ANLN displayed an increasing trend with advancing clinical stage. Furthermore, patients with high ANLN expression levels exhibited poor overall survival rates compared with those with low ANLN expression levels. Subsequent ANLN knockdown experiments indicated elevated cell death rate, and reduced colony formation and migration in both A549 and H1299 cells. Additionally, ANLN knockdown resulted in increased protein expression levels of pyroptosis‑associated molecules, including caspase‑1, NLRP3, cleaved‑GSDMD, IL‑1β, ASC and IL‑18 in both A549 and H1299 cells. In conclusion, ANLN represents an important gene and a promising therapeutic target for LUAD. Its potential as a therapeutic target makes it an interesting candidate for further exploration in the development of novel treatment strategies for LUAD.

摘要

在肺腺癌 (LUAD) 的分子靶向治疗领域已经取得了重大进展。然而,LUAD 的复杂分子模式和高度异质性限制了这些治疗方法在特定患者亚群中的疗效;因此,有必要探索 LUAD 治疗的新靶点。使用基因表达谱交互分析数据库分析 LUAD 中肌动蛋白 (ANLN) 的表达水平。此外,使用 Kaplan-Meier Plotter 评估 ANLN 基因表达与患者生存结局的相关性。随后,通过小干扰 RNA (siRNA) 转染敲低 A549 和 H1299 细胞系中的 ANLN,然后进行 TUNEL、集落形成和 Transwell 测定分别评估细胞死亡、集落形成和迁移。此外,通过 Western blot 分析分析 ANLN 敲低后 caspase-1、白细胞介素 (IL)-18 (IL-18)、IL-1β、NLR 家族富含pyrin 域的 3 (NLRP3)、凋亡相关斑点样蛋白含有 CARD 结构域 (ASC) 和裂解的 gasdermin D (GSDMD) 的表达水平。结果表明,与相邻正常样本相比,LUAD 组织中 ANLN mRNA 表达显著增加。此外,ANLN 的表达水平随着临床分期的进展呈上升趋势。此外,与 ANLN 低表达水平的患者相比,ANLN 高表达水平的患者总体生存率较差。随后的 ANLN 敲低实验表明,A549 和 H1299 细胞的细胞死亡率升高,集落形成和迁移减少。此外,ANLN 敲低导致 A549 和 H1299 细胞中细胞焦亡相关分子的蛋白表达水平升高,包括 caspase-1、NLRP3、cleaved-GSDMD、IL-1β、ASC 和 IL-18。总之,ANLN 是 LUAD 的一个重要基因和有前途的治疗靶点。作为一个治疗靶点,它具有成为 LUAD 新型治疗策略开发的潜在有吸引力的候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3ada/10433705/bfcb8d2a6e60/mmr-28-03-13064-g00.jpg

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