■本研究旨在研究ASAP1(ADP核糖基化因子鸟苷酸激酶1)对胃癌(GC)细胞恶性行为的影响,并阐明参与癌症发生发展的潜在分子机制。
■我们使用CCK8,集落形成,评估ASAP1过表达和敲低对GC细胞恶性肿瘤的影响,流式细胞术(膜联蛋白V/碘化丙啶),Transwell迁移,入侵,和划痕试验。Westernblot分析用于评估ASAP1对血管生成的影响,基质金属蛋白酶(MMPs),凋亡蛋白,上皮间质转化(EMT)相关蛋白,以及AKT和p-AKT。还在携带BGC823细胞来源的肿瘤的裸小鼠中评估了ASAP1敲低的影响。
■我们的研究结果表明,ASAP1在GC细胞中显著过表达,加强它们的扩散,入侵,和移民,同时减少细胞凋亡。相反,ASAP1敲除逆转了这些影响,显着增加裂解的胱天蛋白酶3(Casp3)的表达,PARP,和上皮标记E-cadherin,显著降低MMP2、MMP9、VEGFA、和间充质标志物如N-钙粘蛋白和波形蛋白。此外,它减少了AKT,和p-AKT水平(P<0.01)。ASAP1敲低后,裸鼠的肿瘤生长受到抑制。
■ASAP1的过表达显著促进GC细胞的恶性行为,而它的击倒减少了这些影响。这种调节可能是通过VEGFA的下调,导致血管生成减少,cleaved-Casp3和cleaved-PARP过表达,以及MMPs的减少,EMT,AKT,和p-AKT活性。
UNASSIGNED: This study aims to investigate the influence of ASAP1 (ADP ribosylation factor guanylate kinase 1) on the malignant behavior of gastric cancer (GC) cells and to elucidate the potential molecular mechanisms involved in cancer development and progression.
UNASSIGNED: We assessed the impact of ASAP1 overexpression and knockdown on GC cell malignancy using CCK8, colony formation, flow cytometry (Annexin V/propidium iodide), Transwell migration, invasion, and scratch assays. Western blot analysis was used to assess the effects of ASAP1 on angiogenesis, matrix metalloproteinases (MMPs), apoptotic proteins, epithelial-mesenchymal transition (EMT)-related proteins, as well as AKT and p-AKT. The influence of ASAP1 knockdown was also evaluated in nude mice bearing BGC823 cell-derived tumors.
UNASSIGNED: Our findings revealed that ASAP1 was significantly overexpressed in GC cells, enhancing their proliferation, invasion, and migration, while reducing apoptosis. Conversely, ASAP1 knockdown reversed these effects, markedly increasing the expression of cleaved-caspase 3 (Casp3), PARP, and the epithelial marker E-cadherin, and significantly decreasing MMP2, MMP9, VEGFA, and mesenchymal markers such as N-cadherin and vimentin. Additionally, it reduced AKT, and p-AKT levels (P < 0.01). Tumor growth in nude mice was suppressed following ASAP1 knockdown.
UNASSIGNED: The overexpression of ASAP1 significantly promotes malignant behaviors in GC cells, whereas its knockdown diminishes these effects. This modulation is potentially through the downregulation of VEGFA, leading to reduced angiogenesis, Cleaved-Casp3 and Cleaved-PARP overexpression, and a decrease in MMPs, EMT, AKT, and p-AKT activity.