m6A阅读器IGF2BP2在胰腺癌中的表达及意义

Expression and significance of m6A reader IGF2BP2 in pancreatic cancer

摘要:

背景与目的 N⁶-甲基腺苷（m6A）表观遗传学修饰对基因转录后的表达及生理、病理过程，包括肿瘤发生都有重要的调控作用。m6A阅读器IGF2BP2能够显著增强mRNA的稳定性和翻译效率，并在多种肿瘤中存在表达异常。然而，目前IGF2BP2在胰腺癌中的具体生物学功能尚不明确。因此，本研究探讨m6A阅读器IGF2BP2在胰腺癌中的表达及对胰腺癌细胞功能的影响。方法 采用癌症基因组图谱（TCGA）、基因型及基因表达量关联数据库（GTEX）和基因表达综合数据库（GEO）分析m6A编辑器、擦除器和阅读器相关基因的表达水平，并采用Kaplan-Meier生存曲线分析m6A阅读器IGF2BP2表达与胰腺癌患者预后的关系；采用免疫组化在胰腺癌组织及癌旁组织临床标本中验证IGF2BP2的表达水平；采用CCK-8实验、流式细胞周期检测、平板克隆形成实验、Transwell分析m6A阅读器IGF2BP2敲低后胰腺癌细胞增殖活性、细胞周期、克隆形成数目与迁移能力的变化。结果 TCGA-GTEX及GEO数据库分析显示，m6A阅读器基因IGF2BP2在胰腺癌组织中高表达（均P<0.05），且高表达与不良的总体生存期相关（均P<0.05）。临床标本的免疫组化结果证实m6A阅读器IGF2BP2在胰腺癌组织的表达高于癌旁组织。功能性实验结果显示，IGF2BP2基因敲减后，两株细胞的增殖能力明显减弱，且细胞周期更多停在静止期（G0~G1期），细胞克隆形成数目明显减少，细胞迁移能力明显降低（均P<0.05）。结论 m6A阅读器IGF2BP2在胰腺癌组织中呈高表达，且与胰腺癌患者不良预后密切相关，其作用机制可能与促进癌细胞生长与迁移有关。

Abstract:

Background and Aims N⁶-methyladenosine (m6A) epigenetic modification plays a crucial role in post-transcriptional gene expression regulation and various physiological and pathological processes, including tumorigenesis. The m6A reader IGF2BP2 significantly enhances mRNA stability and translation efficiency and is abnormally expressed in multiple cancers. However, the specific biological function of IGF2BP2 in pancreatic cancer remains unclear. Therefore, this study investigated the expression of the m6A reader IGF2BP2 in pancreatic cancer and its effects on pancreatic cancer cell functions.Methods The expression levels of m6A-related writers, erasers, and readers were analyzed using The Cancer Genome Atlas (TCGA), the Genotype-Tissue Expression (GTEX) database, and the Gene Expression Omnibus (GEO). Kaplan-Meier survival analysis was conducted to assess the relationship between IGF2BP2 expression and the prognosis of pancreatic cancer patients. Immunohistochemistry was used to validate IGF2BP2 expression in clinical specimens of pancreatic cancer tissues and adjacent normal tissues. Functional experiments, including CCK-8 assay, flow cytometry for cell cycle analysis, colony formation assay, and Transwell migration assay, were performed to evaluate changes in cell proliferation, cell cycle distribution, colony formation ability, and migration capacity after IGF2BP2 knockdown in pancreatic cancer cells.Results TCGA-GTEX and GEO database analyses showed that IGF2BP2 was highly expressed in pancreatic cancer tissues (both P<0.05) and that its high expression was associated with poor overall survival (both P<0.05). Immunohistochemical staining of clinical specimens confirmed that IGF2BP2 protein expression was higher in pancreatic cancer than in adjacent normal tissue. Functional experiments demonstrated that IGF2BP2 knockdown significantly reduced the proliferation ability of pancreatic cancer cells, arrested more cells in the G0-G1 phase, decreased colony formation, and impaired cell migration (all P<0.05).Conclusion The m6A reader IGF2BP2 is highly expressed in pancreatic cancer tissues and is closely associated with poor prognosis in patients with this disease. Its mechanism of action may be related to the promotion of cancer cell growth and migration.