EGF自分泌介导EMT对Morris Hepatoma 3924A肝癌细胞的影响
Experimental study on the effect of EGF autocrine mediated EMT on Morris Hepatoma 3924A liver cancer cells
沙天宇 1胡若涵 1王广志 1宁厚法1
作者信息
- 1. 山东第二医科大学医学影像学院,山东 潍坊 261053
- 折叠
摘要
目的 通过观察上皮细胞-间充质转化(EMT)相关蛋白表达变化探讨表皮生长因子(EGF)自分泌对Morris Hepatoma 3924A肝癌细胞的影响.方法 制作Morris Hepatoma 3924A肝癌细胞CoCl2 缺氧模型.通过Western Blot分析EGF自分泌对肿瘤EMT相关蛋白的表达变化,CCK-8 法检测肝细胞癌细胞增殖能力变化,划痕实验检测细胞侵袭能力改变的情况.结果 ①CCK8 实验显示:在轻度缺氧状态下(<150μmol/L CoCl2)肿瘤细胞增殖受限不明显,AG-1478 的48h IC50 为10μmol/L;②Western Blot显示:CoCl2 组的E-cadherin表达减低,而N-cadherin及EGF表达增加(P<0.05),CoCl2+AG-1478 组E-cadherin表达增加,N-cadherin及EGF表达减少(P<0.05);③划痕实验显示:CoCl2 组有更高的迁移能力,人为抑制EGF自分泌后迁移能力明显减弱(P<0.05).结论 肿瘤细胞在缺氧状态下EGF自分泌会对自身起正反馈效果,抑制EGF自分泌可有效抑制肿瘤细胞EMT过程,降低肿瘤细胞的迁移能力.
Abstract
Objective To explore the effect of EGF autocrine on EMT after liver cancer intervention by ob-serving changes in EMT related proteins.Methods A Morris Hepatoma 3924A liver cancer cell model was established by cobalt chloride hypoxia.The expression changes of tumor EMT related proteins caused by EGF autocrine were ana-lyzed by Western blot.The proliferation ability of liver cancer cells under different treatment methods was detected by the CCK-8 method.The changes in cell invasion ability were detected by a scratch assay.Results ①CCK8 experiment showed that under mild hypoxia(<150μmol/L),the proliferation of tumor cells is not significantly restricted with cobalt chloride,and the 48h IC50 of AG-1478 is 10μmol/L;②Western blot showed that compared with the control group,the expression of E-cadherin was reduced in the cobalt chloride group,while the expression of N-cadherin and EGF was in-creased,with statistical significance(P<0.05);The expression of E-cadherin increased,while the expression of N-cad-herin and EGF decreased in the cobalt chloride+AG-1478 group compared to the cobalt chloride group,and the differ-ence was statistically significant(P<0.05);③The scratch test showed that the cobalt chloride group had higher migra-tion ability compared to the normal control group,and the migration ability was significantly reduced after inhibiting EGF autocrine(P<0.05).Conclusion Under hypoxic conditions,tumor cells exhibit a positive feedback effect on their own secretion of EGF;Inhibiting EGF autocrine can effectively inhibit the EMT process of tumor cells;Inhibiting EGF auto-crine can reduce the migration ability of tumor cells.
关键词
表皮生长因子/上皮细胞-间充质转化/肝细胞肝癌Key words
Epidermal growth factor/Epithelial-mesenchymal transition/Hepatocellular carcinoma引用本文复制引用
基金项目
山东省自然科学基金面上项目(ZR2020MH293)
潍坊市科技发展计划项目(医学类)(2022YX050)
出版年
2024
NETL
NSTL
万方数据