| RNA结合蛋白ZC3H15介导非小细胞肺癌发生的机制研究 |
| 投稿时间：2025-05-12 修订日期：2025-06-03 点此下载全文 |
| 引用本文：缪苏南,舒素辉,吕伟,郑婷,张二宝.RNA结合蛋白ZC3H15介导非小细胞肺癌发生的机制研究[J].医学研究杂志,2025,54(10):45-52 |
| DOI：
10.11969/j.issn.1673-548X.2025.10.009 |
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| 基金项目:国家自然科学基金资助项目(面上项目)(82172992) |
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| 中文摘要:目的 探究RNA结合蛋白ZC3H15在非小细胞肺癌中激活的原因及其在非小细胞肺癌发生中的生物学功能。方法 基于癌症基因组图谱(The Cancer Genome Atlas, TCGA)数据库的全基因组测序数据,分析ZC3H15拷贝数变异与其表达的关系,并探究ZC3H15在癌旁正常组织和非小细胞肺癌组织中的差异表达、在不同TNM分期中的差异表达及预后分析。在A549细胞和PC9细胞中,将细胞分为si-NC组和si-c-Myc组,通过实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction, RT-qPCR)法、染色质免疫共沉淀实验探究转录因子c-Myc与ZC3H15之间的调控关系。在A549细胞和PC9细胞中,将细胞分为si-NC组、si-ZC3H15 1#组、si-ZC3H15 2#组、Vector组、pcDNA-ZC3H15组,采用RT-qPCR法检测ZC3H15的表达水平,MTT实验检测细胞活力,克隆形成实验检测细胞增殖能力,Transwell实验检测细胞迁移能力。在A549细胞中稳定转染sh-Ctrl和sh-ZC3H15,采用裸鼠皮下荷瘤实验检测ZC3H15在体内对非小细胞肺癌细胞增殖能力的影响。结果 TCGA非小细胞肺癌数据分析显示,拷贝数扩增组ZC3H15的表达水平高于拷贝数缺失组,且ZC3H15拷贝数与其表达水平呈正相关(P<0.05)；配对和非配对非小细胞肺癌组中ZC3H15的表达水平较正常组均上调(P<0.05)；与Ⅰ/Ⅱ期组比较,Ⅲ/Ⅳ期组的ZC3H15表达水平增加(P<0.05)；ZC3H15低表达组的生存率高于ZC3H15高表达组(P<0.05)。与si-NC组比较,si-c-Myc组ZC3H15的表达水平及c-Myc抗体处理后Input参照百分比均降低(P<0.05)。RT-qPCR法检测结果显示,在A549细胞和PC9细胞中,si-ZC3H15 1#组和si-ZC3H15 2#组的ZC3H15表达水平均低于si-NC组(P<0.05)；pcDNA-ZC3H15组的ZC3H15表达水平高于Vector组(P<0.05)。进一步细胞功能研究发现,敲低ZC3H15非小细胞肺癌细胞增殖及迁移能力均降低(P<0.05),过表达ZC3H15可增强非小细胞肺癌细胞增殖及迁移能力(P<0.05)。在体内水平,裸鼠皮下荷瘤实验表明,ZC3H15敲低可显著降低肿瘤体内生长速率(P<0.05)。结论 拷贝数扩增和c-Myc转录激活是ZC3H15在非小细胞肺癌中激活的重要原因,ZC3H15是促进非小细胞肺癌发生的关键功能分子,可为临床非小细胞肺癌诊疗提供一定的理论依据。 |
| 中文关键词:拷贝数变异 非小细胞肺癌 RNA结合蛋白 ZC3H15 c-Myc |
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| Mechanism of the RNA Binding Protein ZC3H15 in Mediating the Tumorigenesis of Non-small Cell Lung Cancer. |
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| Abstract:Objective To explore the reasons for the activation of the RNA-binding protein ZC3H15 in non-small cell lung cancer and its biological functions in the tumorigenesis of non-small cell lung cancer. Methods Based on the whole genome sequencing data of The Cancer Genome Atlas (TCGA) database, the relationship between ZC3H15 copy number variation and its expression was analyzed, and the differential expression of ZC3H15 between normal and non-small cell lung cancer groups, its variation across different TNM stages, and its prognostic analysis were investigated. A549 cells and PC9 cells were divided into si-NC group and si-c-Myc group, respectively. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and chromatin immunoprecipitation were used to explore the regulatory relationship between the transcription factor c-Myc and ZC3H15. A549 cells and PC9 cells were divided into si-NC group, si-ZC3H15 1# group, si-ZC3H15 2# group, Vector group, pcDNA-ZC3H15group. RT-qPCR was used to detect the expression of ZC3H15, MTT assay was used to detect cell viability, colony formation assay was used to examine cell proliferation capacity, and Transwell assay was used to measure cell migration ability. The A549 cells were stably transfected with sh-Ctrl and sh-ZC3H15, and the nude mouse xenograft model was used to detect the impact of ZC3H15 on the proliferation ability of non-small cell lung cancer cells in vivo. Results TCGA non-small cell lung cancer data analysis showed that the expression level of ZC3H15 in the copy-number-amplified group was higher than that in the copy-number-deleted group (P<0.05). ZC3H15 copy number was positively correlated with expression level (P<0.05). In both paired and unpaired non-small cell lung cancer groups, the expression level of ZC3H15 was up-regulated compared to the normal group (P<0.05). The expression level of ZC3H15 was higher in stage Ⅲ/Ⅳ group than that in stage Ⅰ/Ⅱ group (P<0.05). The low-expression ZC3H15group had a higher survival rate than the high-expression group (P<0.05). Compared to the si-NC group, the expression level of ZC3H15 and Input reference percentage after c-Myc antibody treatment in the si-c-Myc group were both decreased (P<0.05). The results of RT-qPCR showed that in A549 cells and PC9 cells, the expression level of ZC3H15 in the si-ZC3H15 1# and si-ZC3H15 2# groups were lower than those in the si-NC group (P<0.05), while the expression level of ZC3H15 in the pcDNA-ZC3H15group was higher than that in the Vector group (P<0.05). Further cell function studies found that the proliferation and migration capacity of non-small cell lung cancer cells with ZC3H15 knockdown were decreased (P<0.05), and the overexpression of ZC3H15 could enhance the proliferation and migration capacity of non-small cell lung cancer cells (P<0.05). At the in vivo level, subcutaneous tumor-bearing experiments in nude mice showed that ZC3H15 knockdown could significantly reduced the tumor growth rate (P<0.05). Conclusion Copy number amplification and c-Myc transcriptional activation were important reasons for ZC3H15 activation in non-small cell lung cancer. ZC3H15 is a key functional molecule to promote the tumorigenesis of non-small cell lung cancer, and can provide some theoretical basis for clinical diagnosis and treatment of non-small cell lung cancer. |
| keywords:Copy number variation Non-small cell lung cancer RNA-binding protein ZC3H15 c-Myc |
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