Lu Zhou, Huang Xinkun, Shen Qicheng, Chen Erlin, Feng Ying
Department of Gastrointestinal Surgery, Medical School of Nantong University, Affiliated Hospital of Nantong University, Nantong, 226001, China.
Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, 20 Xisi Street, Nantong, 226001, Jiangsu, China.
Biochem Genet. 2024 May 27. doi: 10.1007/s10528-024-10841-2.
Granzyme B (GZMB), a critical member of the Gr gene family, is known to play an essential role in diverse physiological and pathological processes such as inflammation, acute and chronic inflammatory diseases, and cancer progression. In this study, we delve deeper into the role of GZMB within the context of gastric cancer (GC) to examine its expression patterns and functional implications. To accomplish this, we applied a combination of quantitative real-time polymerase chain reaction, western blotting, and immunohistochemistry techniques. These methodologies allowed us to accurately gauge GZMB expression levels in GC tissues and investigate their correlation with various clinical-pathological variables. Our secondary focus was to discern the regulatory influence of GZMB on GC cell biology. We used an array of assays including cell counting kit-8 (CCK-8), colony formation, 5-ethynyl-2'-deoxyuridine, and migration assays. The effect of GZMB on gastric cancer progression was further validated through a subcutaneous xenograft mouse model. Our findings underscored that GZMB mRNA and protein levels were upregulated in GC tissues, a feature that showed a significant correlation with GC staging. We also discovered that a decrease in GZMB expression via knockdown experiments suppressed the proliferation and migration capabilities of GC cells. This effect was manifested through diminished expression levels of epithelial-mesenchymal transition (EMT) markers. In stark contrast, the overexpression of GZMB through plasmid transfection appeared to enhance the proliferation and migration abilities of GC cells. This was coupled with an upregulation in EMT expression. Our study concludes by emphasizing that GZMB promotes the growth, migration, and EMT processes in gastric cancer. In vitro, cell-based experiments and in vivo xenograft mouse models confirm this. Our findings provide a more comprehensive understanding of GZMB's role in gastric cancer pathogenesis, potentially opening doors for novel therapeutic strategies targeting this molecular pathway.
颗粒酶B(GZMB)是Gr基因家族的关键成员,已知在多种生理和病理过程中发挥重要作用,如炎症、急慢性炎症性疾病和癌症进展。在本研究中,我们更深入地探讨了GZMB在胃癌(GC)中的作用,以研究其表达模式和功能意义。为此,我们应用了定量实时聚合酶链反应、蛋白质免疫印迹和免疫组织化学技术的组合。这些方法使我们能够准确测量GC组织中GZMB的表达水平,并研究其与各种临床病理变量的相关性。我们的次要重点是识别GZMB对GC细胞生物学的调节影响。我们使用了一系列检测方法,包括细胞计数试剂盒-8(CCK-8)、集落形成、5-乙炔基-2'-脱氧尿苷和迁移检测。通过皮下异种移植小鼠模型进一步验证了GZMB对胃癌进展的影响。我们的研究结果强调,GC组织中GZMB的mRNA和蛋白质水平上调,这一特征与GC分期显著相关。我们还发现,通过敲低实验降低GZMB表达可抑制GC细胞的增殖和迁移能力。这种作用通过上皮-间质转化(EMT)标志物表达水平的降低得以体现。与此形成鲜明对比的是,通过质粒转染过表达GZMB似乎增强了GC细胞的增殖和迁移能力。这伴随着EMT表达的上调。我们的研究最后强调,GZMB促进胃癌的生长、迁移和EMT过程。体外基于细胞的实验和体内异种移植小鼠模型证实了这一点。我们的研究结果为更全面地理解GZMB在胃癌发病机制中的作用提供了依据,可能为针对这一分子途径的新型治疗策略打开大门。