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| RuvB样AAA ATP酶2通过激活Akt/mTOR信号通路促进肺腺癌细胞增殖、迁移及侵袭 |
| RuvB-like AAA ATPase 2 promotes the proliferation, migration and invasion of Lung adenocarcinoma cells by activating Akt/mTOR signaling pathway |
| 投稿时间：2024-11-12 修订日期：2025-02-06 |
| DOI： |
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| 中文关键词: 肺腺癌 RUVBL2 PI3K-Akt/mTOR 增殖 迁移侵袭 |
| 英文关键词:Lung Adenocarcinoma RUVBL2 PI3K-Akt/mTOR Proliferation Migration and Invasion |
| 基金项目:广东省自然科学(项目编号2023A1515110995) |
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| 中文摘要: |
| 摘 要:[目的] 探讨RuvB样AAA ATP酶2(RuvB-like AAA ATPase 2, RUVBL2)在肺腺癌(lung adenocarcinoma, LUAD)中的表达及与预后的相关性,并进一步研究其对肺腺癌细胞系A549及H1299细胞增殖、迁移侵袭能力的影响及其发挥生物学功能的内在机制。[方法]通过R语言信息技术对癌症基因组图谱(The Cancer Genome Atlas, TCGA)数据库、UALCAN数据库(http://ualcan.path.uab.edu)等分析RUVBL2在肺腺癌及肺组织中的表达及与临床病理学参数间的关系。构建Cox风险回归模型,探讨影响肺腺癌生存率的预后因素。利用基因表达谱交互分析2(gene expression profiling interactive analysis 2, GEPIA2)绘制RUVBL2表达与预后的生存曲线。通过在线网站癌症细胞系百科全书(cancer cell line encyclopedia, CCLE)查询肺癌RUVBL2基因表达量，选择A549及H1299细胞模型进行后续研究。转染siRNA,分为阴性对照组(negative control, NC)组、实验组(siRNA-RUVBL2, siRUVBL2)组。采用细胞计数试剂盒-8(cell counting kit-8, CCK8)法检测细胞增殖能力,采用Transwell、划痕实验检测细胞迁移侵袭能力,并通过R软件对TCGA高通量测序数据中RUVBL2表达相关差异基因进行基于基因组百科全书(kyoto encyclopedia of genes and genomes, KEGG)的基因集富集(gene set enrichment analysis, GSEA)分析寻找RUVBL2可能参与调控的通路,通过蛋白免疫印迹实验(Western blot, WB)验证相关通路。最后，通过10例配对的肺腺癌患者癌与癌旁组织样本的WB实验验证RUVBL2的表达差异。[结果] RUVBL2在肺腺癌中表达显著高于癌旁组织,与肺腺癌患者不良的临床预后相关；RUVBL2可作为肺腺癌预后的独立风险因素；在肺腺癌细胞系A549细胞中沉默RUVBL2后,增殖能力降低,但差异没有统计学意义(P=0.264),而在H1299细胞中沉默RUVBL2后,增殖能力显著下降(P=0.001)；肺腺癌细胞系A549及H1299细胞沉默RUVBL2后,迁移、侵袭能力显著减弱(P<0.001)；在沉默RUVBL2后,磷酸化的蛋白激酶B(phosphor-protein kinase B, p-Akt)及磷酸化的哺乳动物雷帕霉素靶蛋白(phosphor-mammalian target of rapamycin, p-mTOR)蛋白表达显著受到抑制。临床组织样本结果验证了RUVBL2在肺腺癌组织高表达。[结论] RUVBL2在肺腺癌高表达,可作为预测肺腺癌预后的独立危险因素，预示不良的临床预后。RUVBL2可能通过激活Akt/mTOR信号转导通路影响肺腺癌细胞系A549及H1299的增殖、迁移及侵袭。 |
| 英文摘要: |
| Abstract: [Objective] To investigate the expression of RuvB like AAA ATPase 2 (RUVBL2) in lung adenocarcinoma (LUAD) and its correlation with prognosis, as well as further studying its impact on the proliferation, migration and invasion capabilities of lung adenocarcinoma cell lines A549 and H1299, and the underlying mechanisms by which it exerts its biological functions. [Methods] Using R language and bioinformatics tools to analyze the expression and clinicopathological parameters of RUVBL2 in lung adenocarcinoma and lung tissue from databased such as The Cancer Genome Atlas (TCGA) and UALCAN(http://ualcan.path.uab.edu). A Cox risk regression model was constructed to explore the prognostic factors influencing the survival rate of lung adenocarcinoma. Utilizing gene expression profiling interactive analysis 2 (GEPIA2) to plot survival curves for RUVBL2 expression and prognosis. Query the expression level of the RUVBL2 gene in lung cancer through the online cancer cell line encyclopedia (CCLE), and the A549 and H1299 cell models were selected for subsequent studies. Transfected siRNA and divided into a negative control group (NC) and an experimental group (siRNA-RUVBL2, siRUVBL2). We used the cell counting kit-8 (CCK8) assay to detect cell proliferation ability. The cell migration and invasion abilities were detected by Transwell and scratch assay, and the gene set enrichment analysis (GSEA) based on kyoto encyclopedia of genes and genomes (KEGG) was performed on R software to analysis RUVBL2 expression-related differential in TCGA high-throughput sequencing data, and the relevant pathways were validated related by (Western Blot, WB). Finally, the differential expression of RUVBL2 was verified by WB experiments of cancer and para-cancer tissue samples from 10 paired lung adenocarcinoma patients. [Results] RUVBL2 is significantly overexpressed in lung adenocarcinoma compared to adjacent non-cancerous tissue and is associated with poor clinical prognosis in lung adenocarcinoma patients; silencing RUVBL2 in A549 cells reduces proliferative capacity, but the difference is not statistically significant(P=0.264),while in H1299 cells significantly decreases proliferative capacity (P=0.001); silencing RUVBL2 in A549 and H1299 cells significantly impairs migration and invasion capabilities (P<0.001); after silencing RUVBL2,the protein expression of phosphorylated protein kinase B (p-Akt) and phosphorylated mammalian target of rapamycin (p-mTOR) was significantly inhibited. The results of clinical tissue samples validated the high expression of RUVBL2 in lung adenocarcinoma tissues. [Conclusion] RUVBL2 was highly expressed in lung adenocarcinoma, can be used as an independent risk factor for predicting the prognosis of lung adenocarcinoma and RUVBL2 may affect the proliferation, migration and invasion of lung adenocarcinoma cell lines A549 and H1299 by activating the Akt/mTOR signaling pathway. |
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