摘要
目的:观察表皮生长因子(EGF)对人乳腺癌细胞上皮间质转化(EMT)的影响,探讨钙蛋白酶(Calpain)在其中的作用。方法:常规培养雌激素受体(ER)阳性乳腺癌细胞MCF-7和ER阴性MDA-MB-231细胞,EGF诱导EMT并适时加入Calpain抑制剂Calpeptin预处理;划痕实验观察细胞迁移能力,蛋白印迹实验检测细胞FN和E-cadherin蛋白表达。结果:EGF(50μg/L)处理24 h和48 h均显著增加MCF-7和MDA-MB-231细胞迁移(P<0.01);EGF处理24 h、48 h、72 h时均能诱导MCF-7和MDA-MB-231细胞FN表达上调和E-cad表达下调(P<0.01),在48 h时具有最佳诱导效应;Calpeptin(10μmol/L)预处理能显著抑制EGF诱导的MCF-7和MDA-MB-231细胞迁移(P<0.01);48 h时, Calpeptin预处理能显著抑制EGF诱导的MCF-7和MDA-MB-231细胞FN表达上调和E-cad表达下调(P<0.01)。结论:EGF能诱导乳腺癌MCF-7和MDA-MB-231细胞EMT和迁移,该效应可能与Calpain的介导作用相关。
Objective: To observe the effect of epithelial growth factor(EGF) on the epithelia-mesenchymal transition(EMT) in the cells of human breast cancer, and to explore the key role of calpain in the process. Methods: Breast cancer cell MCF-7(ER+) and MDA-MB-231(estrogen receptor, ER-) were used as model cells. The cells were treated with EGF to detect its effect on EMT and calpain inhibitors(calpeptin) was also treated to determine the effect of calpain in the process. Cell wound scratch assay was used to detect cell migration ability;Western blotting assay was used to detected the protein expression changes of FN and E-cad in the cells. Results: After treatment with EGF(50 μg/L) for 24 h and 48 h, the migration of MCF-7 and MDA-MB-231 cells increased significantly(P<0.01). EGF treatment induced up-regulation of FN and down-regulation of E-cad expression in MCF-7 and MDA-MB-231 cells for 24 h, 48 h and 72 h(P<0.01). The best induction effect occurred in 48 h. Calpeptin(10 μmol/L) pretreatment significantly inhibited EGF-induced MCF-7 and MDA-MB-231 cell migration(P<0.01). In 48 h, calpeptin pretreatment significantly inhibited EGF-induced up-regulation of FN and down-regulation of E-cad expression in MCF-7 and MDA-MB-231 cells(P<0.01). Conclusion: EGF can induce EMT and migration in breast cancer MCF-7 and MDA-MB-231 cells, and calpain may play a key role in the treatment of breast cancer.
作者
金爱
王宏键
王旭东
JIN Ai;WANG Hongjian;WANG Xudong(Department of Physiology, School of Basic Medicine, Guizhou Medical University, Guiyang 550025, Guizhou, China)
出处
《贵州医科大学学报》
CAS
2019年第4期412-417,共6页
Journal of Guizhou Medical University
基金
国家自然科学基金资助项目(31660345
31360252)
关键词
表皮生长因子
钙蛋白酶
乳腺肿瘤
上皮-间质转化
迁移
epithelial growth factor
calpain
breast cancer
epithelial-mesenchymal transition
migration
