Yao Han-Hui, Zhao Ya-Jun, He Yi-Fu, Huang Da-Bing, Wang Wei
1Department of General Surgery, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230001 P.R. China.
2Department of Medical Oncology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, No. 17 Lujiang Road, Hefei, Anhui, 230001 P.R. China.
Cancer Cell Int. 2019 Feb 27;19:41. doi: 10.1186/s12935-019-0765-6. eCollection 2019.
Angiogenic factor with G-patch and FHA domain 1 (AGGF1), as a newly identified human angiogenic factor, is overexpressed in some types of malignant tumors and closely associated with patient's prognosis. However, the mechanisms involved in the regulation of AGGF1 in gastric cancer (GC) still remain unclear.
In this study, AGGF1 level in GC tissues and cell lines was analyzed by western blot and quantitative real-time polymerase chain reaction (qRT-PCR). After knockdown of AGGF expression by RNA interference in GC cell lines MKN-45 and MGC-803, wound healing and transwell assays were conducted to examine the effects of AGGF1 on migration and invasion. Tumor growth was assessed in a mouse xenograft model in vivo. Furthermore, expression levels of epithelial-mesenchymal transition (EMT) biomarkers and involvement of the Wnt/β-catenin pathway were detected by western blot and qRT-PCR.
Compared to those in normal groups, the protein and mRNA of AGGF1 expression levels were significantly higher both in GC tissues and cell lines (all P < 0.05). Knockdown of AGGF1 dramatically inhibited the invasion and migration of MKN-45 and MGC-803 cells (all P < 0.01) in vitro, and suppressed the tumor growth of nude mice xenograft model in vivo. Western blot revealed alterations in EMT biomarkers, suggesting the role of AGGF1 in EMT. Moreover, we found that downregulated expression of AGGF1 attenuated Wnt/β-catenin related protein expression.
Collectively, knockdown of AGGF1 inhibits the invasion and migration of gastric cancer via epithelial-mesenchymal transition through Wnt/β-catenin pathway.
含G-结构域和FHA结构域的血管生成因子1（AGGF1）作为一种新发现的人类血管生成因子，在某些类型的恶性肿瘤中过表达，且与患者预后密切相关。然而，AGGF1在胃癌（GC）中的调控机制仍不清楚。
在本研究中，通过蛋白质免疫印迹法和定量实时聚合酶链反应（qRT-PCR）分析GC组织和细胞系中AGGF1的水平。在GC细胞系MKN-45和MGC-803中通过RNA干扰敲低AGGF表达后，进行伤口愈合和Transwell实验以检测AGGF1对迁移和侵袭的影响。在体内小鼠异种移植模型中评估肿瘤生长。此外，通过蛋白质免疫印迹法和qRT-PCR检测上皮-间质转化（EMT）生物标志物的表达水平以及Wnt/β-连环蛋白信号通路的参与情况。
与正常组相比，GC组织和细胞系中AGGF1的蛋白质和mRNA表达水平均显著更高（均P < 0.05）。敲低AGGF1显著抑制了MKN-45和MGC-803细胞在体外的侵袭和迁移（均P < 0.01），并抑制了体内裸鼠异种移植模型的肿瘤生长。蛋白质免疫印迹法显示EMT生物标志物发生改变，提示AGGF1在EMT中的作用。此外，我们发现AGGF1表达下调减弱了Wnt/β-连环蛋白相关蛋白的表达。
总体而言，敲低AGGF1通过Wnt/β-连环蛋白信号通路介导的上皮-间质转化抑制胃癌的侵袭和迁移。
