南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (11): 1638-1645.doi: 10.12122/j.issn.1673-4254.2022.11.07

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MiR-4772通过调控卵巢癌免疫相关基因改变肿瘤免疫微环境

赵爱月，苏云霞，傅德强   

  1. 福建医科大学附属第二医院肿瘤内科，福建 泉州 362000
  • 出版日期:2022-11-20 发布日期:2022-11-30

MiR-4772 modulates tumor immune microenvironment by regulating immune-related genes in ovarian cancer

ZHAO Aiyue, SU Yunxia, FU Deqiang   

  1. Department of Oncology, Second Affiliated Hospital of Fujian Medical University, Quanzhou 362000, China
  • Online:2022-11-20 Published:2022-11-30

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摘要： 目的 运用生物信息学分析miR-4772对卵巢癌肿瘤免疫微环境的影响，并深入挖掘其潜在作用机制。方法 基于TCGA数据库的294例卵巢癌患者数据，计算并获得肿瘤组织PD-L1表达最佳阈值；筛选PD-L1高、低表达组之间的差异表达基因（DEG），采用相关性分析获得重要DEG，运用WGCNA分析从DEG筛选出权重基因和PD-L1相关miRNA，重要DEG和权重基因交集获得核心基因；通过ssGSEA分析探讨PD-L1表达和miR-4772表达对卵巢癌肿瘤免疫格局的影响；KEGG法分析核心基因所涉及的信号通路；PPI网络分析核心基因相互作用；采用生存分析确定生存相关的核心基因。采用GEO数据库的399例卵巢癌患者数据，结合miR-4772模拟物和抑制物转染SKOV3细胞并进行基因表达谱测序验证。结果 卵巢癌肿瘤组织PD-L1表达最佳阈值1.31582，即90%分位数；基于PD-L1高、低表达组，筛选出840个差异表达基因，包括549个基因显著上调，291个基因显著下调，20个重要DEG与miR-4772表达密切相关；WGCNA分析筛选48个miR-4772表达相关的权重基因；12个核心基因共存于重要DEG和权重基因。ssGSEA分析显示，PD-L1和miR-4772高表达组展现更活跃的肿瘤组织免疫浸润和功能活性。KEGG功能分析结果显示12个核心基因主要涉及免疫相关信号通路，PPI分析显示12个核心基因存在密切相互作用，CD96和TBX21表达水平与卵巢癌患者生存预后密切相关。SKOV3细胞转染miR-4772模拟物和抑制物后，基因表达谱测序结果显示12个核心基因和PD-L1表达水平随着miR-4772表达水平变化出现相应改变。结论 miR-4772通过作用于12个核心基因，对卵巢癌肿瘤免疫微环境结构组成发挥调控作用。

关键词: 卵巢癌；肿瘤免疫微环境；miR-4772；PD-L1表达最佳阈值

Abstract: Objective To explore the regulatory role of miR-4772 in the formation of tumor immune microenvironment in ovarian cancer. Methods The optimal cutoff level of PD-L1 expression was calculated based on data from 294 ovarian cancer patients in the TCGA database. The differentially expressed genes (DEGs) between high and low PD-L1 expression groups were screened, and the important DEGs were identified by correlation analysis. WGCNA analysis was performed to select the weighted genes and PD-L1-related miRNAs, from which the hub genes were obtained by intersection analysis. ssGSEA analysis was used to evaluate the effect of PD-L1 and miR-4772 expressions on the tumor immune microenvironment in ovarian cancer. KEGG analysis was used to identify the involved signal pathways, and the interactions between the hub genes were mapped by protein-protein interaction (PPI) analysis. Survival analysis was carried out to identify the survival-related hub genes, and the results were validated using the data of 399 patients with ovarian cancer from GEO database and the sequencing results of SKOV3 cells transfected with miR-4772 mimics or inhibitor. Results According the optimal cutoff level of PD-L1 expression of 1.31582 (90th quantile), the patients were divided into high- and low-PD-L1 expression groups. A total of 840 DEGs were identified, including 549 significantly up-regulated genes and 291 down- regulated genes. Among them, 20 important DEGs were found to closely correlate with miR-4772 expression, and WGCNA analysis identified 48 weighted genes significantly correlated with miR-4772. Twelve genes were identified as both key DEGs and weighted genes and were treated as the hub genes. ssGSEA analysis showed that both the patients with high PD-L1 expressions and those with high miR-4772 expressions showed more active immune infiltration and functional activity. The 12 hub genes were involved mainly in immune- related signaling pathways, and PPI analysis suggested significant interactions among the hub genes. The two hub genes CD96 and TBX21 showed close correlation with the survival of ovarian cancer patients. The sequencing results of SKOV3 cells transfected with miR-4772 mimics or inhibitor showed that the changes in miR-4772 expression level caused obvious changes in the expressions of the 12 hub genes and PD-L1. Conclusion MiR-4772 plays a regulatory role in the formation of tumor immune microenvironment in ovarian cancer by regulating 12 hub genes.

Key words: ovarian cancer; tumor immune microenvironment; miR-4772; optimal cutoff point of PD-L1 expression