| 中文关键词: 肺腺癌 RuvB样AAA ATP酶2 AKT/mTOR 增殖 迁移侵袭 |
| 英文关键词:lung adenocarcinoma RuvB-like AAA ATPase 2 AKT/mTOR proliferation migration and invasion |
| 基金项目:广东省自然科学基金项目（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、实验组（siRNA-RUVBL2，si-RUVBL2）。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。通过10例配对的肺腺癌患者癌组织与癌旁组织样本的WB实验验证RUVBL2表达差异。［结果］ RUVBL2在肺腺癌中表达显著性高于癌旁组织，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，但差异无统计学意义（P=0.264）；而在H1299细胞中沉默RUVBL2后，增殖能力显著性下降（P=0.001）。肺腺癌细胞系A549及H1299细胞沉默RUVBL2后，迁移、侵袭能力显著性减弱（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）蛋白表达受到显著性抑制。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 effect of RuvB-like AAA ATPase 2 (RUVBL2) on proliferation， migration and invasion capabilities of lung adenocarcinoma cells and the underlying mechanisms. A549 and H1299， by which it exerts its biological functions. ［Methods］ The relationship between RUVBL2 expression and clinicopathological parameters of lung adenocarcinoma was analyzed using R language and bioinformatics tools based on the data from 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. The gene expression profiling interactive analysis 2 (GEPIA2) was used to plot survival curves of lung cancer patients for RUVBL2 expression. The expression level of the RUVBL2 gene in lung cancer was obtained through the online cancer cell line encyclopedia (CCLE). Lung adenocarcinoma A549 and H1299 cells were transfected with siRNA-RUVBL2. The cell proliferation ability was detected by cell counting kit-8 (CCK8) assay， the cell migration and invasion abilities were detected by transwell and scratch assay. The differentially expressed genes related to RUVBL2 were obtained from TCGA high-throughput sequencing data， the gene set enrichment analysis(GSEA) was performed with R software based on Kyoto encyclopedia of genes and genomes (KEGG) to find the relevant RUVBL2 pathways. Finally， the differential expression of RUVBL2 was verified by Western blot in 10 paired of lung adenocarcinoma and pericancerous tissue samples. ［Results］ RUVBL2 was significantly overexpressed in lung adenocarcinoma compared to adjacent non-cancerous tissue， and was associated with poor clinical prognosis of patients. Silencing RUVBL2 in H1299 cells significantly decreased cell proliferative capacity (P=0.001)， but not in A549 cells (P=0.264). Silencing RUVBL2 in A549 and H1299 cells significantly inhibited cell migration and invasion capabilities (P<0.001). After silencing RUVBL2， the 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 is highly expressed in lung adenocarcinoma， which is associated with unfavorable prognosis of patients. RUVBL2 may affect the proliferation， migration and invasion of lung adenocarcinoma cells by activating the AKT/mTOR signaling pathway. |
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