Department of Traditional Chinese Medicine, People's Hospital of Longhua, Shenzhen, Guangdong 518109, China.
Science and Technology Innovation Center, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 518100, China.
J Proteome Res. 2024 Oct 4;23(10):4579-4588. doi: 10.1021/acs.jproteome.4c00472. Epub 2024 Sep 22.
This study aimed to investigate the dysregulated proteins and the underlying mechanisms of gastric precancerous lesions. Proteomic and phosphoproteomic methods were used to characterize the proteome and phosphoproteome profiles of -methyl--nitro--nitrosoguanidine (MNNG)-induced gastric precancerous lesions. The hub differentially expressed proteins (DEPs) and phosphoproteins (DEPPs) were identified by using differential expression and protein-protein interaction network analyses. Western blot assay, quantitative reverse transcription (qRT)-PCR, and CCK-8 assays detected the expression of Rps3, -cadherin, -cadherin, AKT, -AKT, and β-catenin and verified the roles of Rps3 on the MNNG-induced human gastric epithelial cell line (GES-1) cells. Hub DEPs and phosphoproteins Rps3, Akt1, and Ctnnb1 were significantly correlated with five dendritic cells (DCs) pathways, and Akt1 and Ctnnb1 were significantly negatively correlated with Rps3. MNNG administration markedly reduced the Rps3 mRNA and protein expression levels (all < 0.05). Overexpression of Rps3 significantly inhibited tumorigenesis of MNNG-induced GES-1 cells (all < 0.01) and changed the protein levels of -cadherin, -cadherin, AKT, -AKT, and β-catenin. Similarly, SC79 treatment substantially increased the expression of interleukin (-6, -10, and vascular endothelial growth factor (all < 0.05). Rps3 was poorly expressed in precancerous gastric lesions. Correspondingly, overexpression of Rps3 promoted DC maturation via the AKT/β-catenin pathway, inhibiting the progression of gastric precancerous lesions.
本研究旨在探讨胃癌前病变中失调蛋白及其潜在机制。采用蛋白质组学和磷酸蛋白质组学方法，对甲基亚硝胍（MNNG）诱导的胃癌前病变的蛋白质组和磷酸蛋白质组图谱进行了特征分析。通过差异表达和蛋白质相互作用网络分析，鉴定了枢纽差异表达蛋白（DEPs）和磷酸蛋白（DEPPs）。Western blot 检测、实时定量逆转录（qRT）-PCR 和 CCK-8 检测 Rps3、E-钙黏蛋白、N-钙黏蛋白、AKT、p-AKT 和 β-连环蛋白的表达，并验证了 Rps3 在 MNNG 诱导的人胃上皮细胞系（GES-1）细胞中的作用。枢纽 DEPs 和磷酸蛋白 Rps3、Akt1 和 Ctnnb1 与五个树突状细胞（DC）途径显著相关，Akt1 和 Ctnnb1 与 Rps3 显著负相关。MNNG 给药显著降低了 Rps3 的 mRNA 和蛋白表达水平（均<0.05）。Rps3 的过表达显著抑制了 MNNG 诱导的 GES-1 细胞的肿瘤发生（均<0.01），并改变了 E-钙黏蛋白、N-钙黏蛋白、AKT、p-AKT 和 β-连环蛋白的蛋白水平。同样，SC79 处理显著增加了白细胞介素（-6、-10 和血管内皮生长因子（均<0.05）的表达。Rps3 在癌前胃病变中表达不佳。相应地，Rps3 的过表达通过 AKT/β-连环蛋白通路促进 DC 成熟，抑制胃癌前病变的进展。
