Departments of Hematology-Oncology, College of Medicine, Yeungnam University, Daegu, Republic of Korea.
Departments of Hematology-Oncology, College of Medicine, Yeungnam University, Daegu, Republic of Korea
Anticancer Res. 2024 May;44(5):1973-1981. doi: 10.21873/anticanres.17000.
BACKGROUND/AIM: A role for cold-shock domain (CSD) proteins in abnormal cell proliferation has been suggested in the literature. The aim of this study was to investigate the effect of hepatocyte growth factor (HGF)-induced up-regulation of CSD protein A (CSDA) expression on vascular endothelial growth factor (VEGF) expression and its role in gastric cancer cell invasion and proliferation.
We assessed effects on two gastric cancer cell lines using reverse transcription-polymerase chain reaction, western blotting, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, and CSDA knockdown with short hairpin RNA.
Hepatocyte growth factor (HGF) elevates CSDA levels in gastric cancer cell lines. To elucidate the mechanism by which HGF prompts CSDA expression and its impact on vascular endothelial growth factor (VEGF), we applied the Mitogen Activated Protein Kinase (MAPK) inhibitor PD098059 and conducted analyses using western blot. Following the administration of PD098059, a reduction in the protein levels of HGF-stimulated VEGF was observed. Additionally, silencing of CSDA resulted in diminished levels of both VEGF and phosphorylated extracellular signal-regulated kinase (ERK). The suppression of CSDA also led to reduced HGF-induced cell proliferation and diminished invasive capabilities in vitro. Furthermore, our research pinpointed a potential activator protein-1 (AP-1) binding site within the VEGF promoter zone, validating its activity via chromatin immunoprecipitation assays. Electrophoretic mobility shift assays further disclosed that HGF-induced CSDA augmentation correlates with an increase in AP-1 binding to VEGF.
CSDA is crucial for the proliferation of gastric cancer cells, and the inhibition of this protein could impede the advancement of gastric cancer.
背景/目的：文献中提出冷休克结构域（CSD）蛋白在异常细胞增殖中起作用。本研究旨在探讨肝细胞生长因子（HGF）诱导的 CSD 蛋白 A（CSDA）表达上调对血管内皮生长因子（VEGF）表达的影响及其在胃癌细胞侵袭和增殖中的作用。
我们使用逆转录-聚合酶链反应、western blot、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定和短发夹 RNA 敲低评估了两种胃癌细胞系的影响。
HGF 可提高胃癌细胞系中的 CSDA 水平。为了阐明 HGF 促使 CSDA 表达的机制及其对血管内皮生长因子（VEGF）的影响，我们应用丝裂原活化蛋白激酶（MAPK）抑制剂 PD098059 并进行了 western blot 分析。给予 PD098059 后，观察到 HGF 刺激的 VEGF 蛋白水平降低。此外，CSDA 沉默导致 VEGF 和磷酸化细胞外信号调节激酶（ERK）水平降低。CSDA 的抑制也导致 HGF 诱导的细胞增殖减少和体外侵袭能力降低。此外，我们的研究还在 VEGF 启动子区域内确定了一个潜在的激活蛋白-1（AP-1）结合位点，通过染色质免疫沉淀测定验证了其活性。电泳迁移率变动分析进一步揭示，HGF 诱导的 CSDA 增加与 AP-1 与 VEGF 的结合增加相关。
CSDA 对胃癌细胞的增殖至关重要，抑制这种蛋白可能会阻碍胃癌的进展。
