Fang Jun, Zhang Qiong, Wang Qingrui
Department of Gastrointestinal Surgery, The Affiliated Hospital of Jianghan University, WuHan City No.6 Hospital, Wuhan, 430015, China.
Department of Oncology, The Affiliated Hospital of Jianghan University, WuHan City No.6 Hospital, Wuhan, 430015, China.
Biochem Genet. 2024 Dec 27. doi: 10.1007/s10528-024-11008-9.
Gastric cancer is associated with high morbidity and mortality rates and seriously threatens human life. Our research aimed to explore the effects of poly (A) binding protein cytoplasmic 1 (PABPC1) on gastric cancer cells and elucidate the underlying mechanisms.
PABPC1 levels in gastric cancer cell lines were assessed by western blotting and RT-qPCR. Cell viability, apoptosis, invasion, and migration were analyzed using MTT assay, flow cytometry, wound healing assay, and transwell assay, respectively. The levels of apoptosis-related proteins (caspase 3 and cleaved-caspase 3) were determined using the caspase 3 vitality test kit and western blotting. The levels of epithelial-to-mesenchymal transition-related genes (E-cadherin and N-cadherin) in AGS and MGC803 cells were analyzed using western blotting and RT-qPCR. The phosphatidylinositol 3-kinase (PI3K)/AKT pathway was examined using western blot analysis.
PABPC1 expression was enhanced in gastric cancer cells, especially in AGS and MGC803 cells. Our findings indicate that PABPC1 knockdown by siRNA inhibited PABPC1 expression, repressed gastric cancer cell growth, promoted apoptosis, and enhanced cleaved-caspase 3 expression. Functional assays revealed that PABPC1-siRNA blocked the migration and invasion of gastric cancer cells, dramatically promoted E-cadherin expression, and reduced N-cadherin levels. We also found decreased p-PI3K and p-AKT expression, along with decreased p-PI3K/PI3K and p-AKT/AKT in PABPC1-siRNA-treated gastric cancer cells.
PABPC1 silencing in gastric cancer cells inhibited cell proliferation, metastasis, and epithelial-to-mesenchymal transition, partly by repressing the PI3K/AKT signaling pathway activation. This may provide a theoretical basis for gastric cancer therapeutics.
胃癌具有较高的发病率和死亡率，严重威胁人类生命。我们的研究旨在探讨多聚腺苷酸结合蛋白胞质1（PABPC1）对胃癌细胞的影响，并阐明其潜在机制。
通过蛋白质免疫印迹法和逆转录定量聚合酶链反应（RT-qPCR）评估胃癌细胞系中PABPC1的水平。分别使用MTT法、流式细胞术、伤口愈合试验和Transwell试验分析细胞活力、凋亡、侵袭和迁移情况。使用半胱天冬酶3活力检测试剂盒和蛋白质免疫印迹法测定凋亡相关蛋白（半胱天冬酶3和裂解的半胱天冬酶3）的水平。通过蛋白质免疫印迹法和RT-qPCR分析AGS和MGC803细胞中上皮-间质转化相关基因（E-钙黏蛋白和N-钙黏蛋白）的水平。使用蛋白质免疫印迹分析检测磷脂酰肌醇3-激酶（PI3K）/AKT信号通路。
PABPC1在胃癌细胞中表达增强，尤其是在AGS和MGC803细胞中。我们的研究结果表明，小干扰RNA（siRNA）敲低PABPC1可抑制PABPC1表达，抑制胃癌细胞生长，促进凋亡，并增强裂解的半胱天冬酶3表达。功能试验表明，PABPC1-siRNA可阻断胃癌细胞的迁移和侵袭，显著促进E-钙黏蛋白表达，并降低N-钙黏蛋白水平。我们还发现，在PABPC1-siRNA处理的胃癌细胞中，磷酸化PI3K（p-PI3K）和磷酸化AKT（p-AKT）表达降低，同时p-PI3K/PI3K和p-AKT/AKT也降低。
胃癌细胞中PABPC1沉默可抑制细胞增殖、转移和上皮-间质转化，部分原因是抑制PI3K/AKT信号通路激活。这可能为胃癌治疗提供理论依据。
