| 摘要: |
| ［摘要］ 目的 分析表皮生长因子样结构域9（EGFL9）对表皮生长因子受体（EGFR）/磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（AKT）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路的影响，探讨EGFL9对肝癌细胞增殖、侵袭、迁移的分子机制。方法 通过慢病毒介导的小干扰RNA（siRNA）技术构建针对两个靶点敲减EGFL9表达的Huh-7细胞。选择该研究团队前期构建的EGFL9过表达的Huh-7细胞及EGFL9敲减的JHH-7细胞裸鼠皮下种植瘤进行实验。应用EGFR抑制剂GW2974和AKT抑制剂MK2206处理EGFL9过表达的Huh-7细胞。通过MTT实验评估细胞的增殖活力，通过Transwell实验评估细胞的迁移与侵袭能力。通过流式细胞术检测细胞凋亡情况。通过Western blot实验检测EGFR/PI3K/AKT/mTOR信号通路相关蛋白表达水平。结果 敲减EGFL9的Huh-7细胞的增殖、侵袭、迁移能力均受显著抑制（P<0.05），细胞凋亡率升高（P<0.05）。Western blot实验结果显示，敲减EGFL9可显著抑制肝癌细胞及相应种植瘤瘤体中磷酸化表皮生长因子受体（p-EGFR）、磷酸化磷脂酰肌醇3-激酶（p-PI3K）、磷酸化蛋白激酶B（p-AKT）、磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）的蛋白表达水平（P<0.05），而EGFL9过表达的肝癌细胞中上述蛋白表达水平升高（P<0.05）。经EGFR及AKT抑制剂处理后，EGFL9过表达肝癌细胞相关通路蛋白的磷酸化蛋白水平显著下调（P<0.05），细胞迁移能力减弱（P<0.05）。结论 EGFL9可能通过EGFR/PI3K/AKT/mTOR信号通路促进肝癌细胞的增殖、侵袭及迁移，具有作为肝癌治疗靶点的前景。 |
| 关键词: 表皮生长因子样结构域9 肝细胞癌 分子机制 增殖 侵袭 迁移 |
| DOI：10.3969/j.issn.1674-3806.2026.06.08 |
| 分类号: |
| 基金项目:国家自然科学基金项目（编号：81974442）；广东省自然科学基金项目（编号：2020A1515010799）；广州市科技计划-市校（院）企联合资助项目（编号：2024A03J0670） |
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| EGFL9 enhances the proliferative, invasive and migratory capacities of hepatocellular carcinoma cells via activation of the EGFR/PI3K/AKT/mTOR signaling pathway |
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FENG Jingyang, SHEN Xiaobao, LI Ziye, WANG Bailin, Wu Fan
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Department of Hepatobiliary, Pancreatic, Splenic and Thyroid Surgery, Guangzhou Red Cross Hospital of Jinan University, Guangzhou 510220, China
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| Abstract: |
| ［Abstract］ Objective To analyze the effects of epidermal growth factor-like domain 9(EGFL9) on the EGFR/PI3K/AKT/mTOR signaling pathway and to elucidate the molecular mechanisms by which EGFL9 regulates hepatocellular carcinoma(HCC) cell proliferation, invasion and migration. Methods Huh-7 cells with EGFL9 knockdown targeting two distinct sites were constructed using lentivirus-mediated small interfering RNA(siRNA) technology. The EGFL9-overexpressing Huh-7 cell and EGFL9-knockdown JHH-7 cell-derived subcutaneously implanted tumors in nude mice constructed by the research team in the previous study were selected for experiments. The EGFL9-overexpressing Huh-7 cells were treated with the EGFR inhibitor GW2974 and the AKT inhibitor MK2206. Cell proliferation was assessed by MTT assay. Cell migratory and invasive capacities were assessed by Transwell assay, and cell apoptosis was detected by flow cytometry. The expression levels of EGFR/PI3K/AKT/mTOR signaling pathway-related proteins were detected by Western blot assay. Results The proliferative, invasive and migratory capacities of Huh-7 cells with EGFL9 knockdown were significantly inhibited, and their apoptosis rate was increased. The results of Western blot assay showed that EGFL9 knockdown could significantly suppress the expression levels of phosphorylated epidermal growth factor receptor(p-EGFR), phosphorylated phosphatidylinositol 3-kinase(p-PI3K), phosphorylated protein kinase B(p-AKT), and phosphorylated mammalian target of rapamycin(p-mTOR) in both HCC cells and the corresponding xenograft tumors, whereas the expression levels of these phosphorylated proteins increased in the EGFL9-overexpreesing HCC cells(P<0.05). After treatments with EGFR and AKT inhibitors, the phosphorylated protein levels of the pathway proteins related to EGFL9-overexpressing HCC cells were significantly down-regulated(P<0.05), and the cell migratory capacity was weakened. Conclusion EGFL9 may enhance the proliferative, invasive and migratory capacities of HCC cells via the EGFR/PI3K/AKT/mTOR signaling pathway, and has the potential to serve as a therapeutic target for HCC. |
| Key words: Epidermal growth factor-like domain 9(EGFL9) Hepatocellular carcinoma(HCC) Molecular mechanisms Proliferation Invasion Migration |
