Ning Weiwei, Yang Qingxu, Li Zhengbiao, Xie Ming
Department of General Surgery, Digestive Disease Hospital, Affiliated Hospital of Zunyi Medical University, China.
Department of General Surgery, Digestive Disease Hospital, Affiliated Hospital of Zunyi Medical University, China.
Mol Cell Probes. 2024 Dec;78:101990. doi: 10.1016/j.mcp.2024.101990. Epub 2024 Nov 25.
In gastric cancer (GC), tumor cell metastasis to lymph node may occur, and can be impacted by synaptojanin 2 (SYNJ2). Herein, we explored the mechanism of SYNJ2 in the progress of GC. SYNJ2 level in GC tissues was predicted by GEPIA database. After GC cells were transfected with short hairpin RNA against SYNJ2 (shSYNJ2), shGRB2, SYNJ2 overexpression plasmid and growth factor receptor-bound protein 2 (GRB2) overexpression plasmid, the mRNA levels of SYNJ2 and GRB2 in GC cells were quantified by qRT-PCR. CCK-8, flow cytometry, wound healing, transwell and tube formation assays were performed for detecting viability, apoptosis, migration, invasion and angiogenesis of GC cells. Protein levels of GRB2, vascular endothelial growth factor (VEGF), E-Cadherin, N-Cadherin and Vimentin in GC cells were measured by Western blot. The relationship between SYNJ2 and GRB2 was assessed by Co-immunoprecipitation (CO-IP) assay. SYNJ2 was highly expressed in GC tissues and cells. SYNJ2 overexpression promoted viability, migration, invasion, angiogenesis and GRB2 level, and inhibited apoptosis of GC cells, while shSYNJ2 exhibited opposite effects. GRB2 overexpression boosted yet shGRB2 suppressed cell migration, invasion and angiogenesis. Notably, SYNJ2 could interact with GRB2. GRB2 overexpression and shGRB2 reversed the effects of shSYNJ2 and overexpressed SYNJ2 on cell migration, invasion and angiogenesis and levels of metastasis-related proteins, respectively. In conclusion, SYNJ2 promotes GC cell metastasis and angiogenesis by up-regulating GRB2.
在胃癌（GC）中，肿瘤细胞可能会发生向淋巴结的转移，并且这一过程可能受突触结合蛋白2（SYNJ2）的影响。在此，我们探究了SYNJ2在胃癌进展中的作用机制。通过GEPIA数据库预测胃癌组织中SYNJ2的水平。在用针对SYNJ2的短发夹RNA（shSYNJ2）、shGRB2、SYNJ2过表达质粒以及生长因子受体结合蛋白2（GRB2）过表达质粒转染胃癌细胞后，采用qRT-PCR定量检测胃癌细胞中SYNJ2和GRB2的mRNA水平。进行CCK-8、流式细胞术、伤口愈合、Transwell和管腔形成实验，以检测胃癌细胞的活力、凋亡、迁移、侵袭和血管生成情况。通过蛋白质免疫印迹法检测胃癌细胞中GRB2、血管内皮生长因子（VEGF）、E-钙黏蛋白、N-钙黏蛋白和波形蛋白的蛋白水平。通过免疫共沉淀（CO-IP）实验评估SYNJ2与GRB2之间的关系。SYNJ2在胃癌组织和细胞中高表达。SYNJ2过表达促进了胃癌细胞的活力、迁移、侵袭、血管生成以及GRB2水平，并抑制了胃癌细胞的凋亡，而shSYNJ2则表现出相反的作用。GRB2过表达促进了细胞迁移、侵袭和血管生成，而shGRB2则抑制了这些过程。值得注意的是，SYNJ2可与GRB2相互作用。GRB2过表达和shGRB2分别逆转了shSYNJ2和过表达SYNJ2对细胞迁移、侵袭和血管生成以及转移相关蛋白水平的影响。总之，SYNJ2通过上调GRB2促进胃癌细胞转移和血管生成。
