摘要
目的筛选能抑制BRAF^(V600E) CT26细胞株增殖的中药单体化合物。方法通过慢病毒载体感染,构建BRAF^(V600E)稳定表达的CT26细胞稳转株。MTT检测细胞增殖,划痕实验检测细胞迁移,蛋白印迹检测MEK/ERK信号通路蛋白表达。Discovery Studio筛选BRAF^(V600E)高亲和中药单体化合物,并用MTT验证化合物抑制细胞增殖效果。结果与野生型CT26细胞相比,BRAF^(V600E) CT26细胞的增殖和迁移能力明显增强,MEK/ERK通路被进一步激活。芦荟素(aloin)、安格洛苷C(angoroside C)、杯苋甾酮(cyasterone)对BRAFV600E CT26细胞的抑制效果优于野生型CT26细胞(P <0. 05),并能明显降低BRAFV600E的蛋白表达。结论芦荟素、安格洛苷C、杯苋甾酮可能是结肠癌BRAFV600E突变的天然抑制剂。
Aim To screen BRAF^V600E CT26 cell inhibitors from monomers of traditional Chinese medicine (TCM).Methods CT26 cell line was constructed with lentivirus plasmid to stably express BRAF^V600E .The proliferation,migration and expression of related proteins in MEK/ERK signaling pathway were detected.The monomers of TCM were detected for biological activities as potential BRAF^V600E inhibitors by Discovery Studio 4.0,and further evaluated by MTT assay.Results The proliferation and migration of BRAF^V600E CT26 cells were obviously strengthened compared with wild type control.The expressions of proteins in MEK/ERK pathway were also activated in BRAF^V600E CT26 cells.Compared with wild type control,Aloin,Angoroside C and Cyasterone exhibited the potent effect against BRAF^V600E in CT26 cells ( P <0.05),and could down-regulate the expression of BRAF^V600E .Conclusion Aloin,Angoroside C,Cyasterone might be the potent inhibitors against BRAF^V600E for colon treatment.
作者
张欢欢
方杰
毛诗莹
杨扬
谢珲
余陈欢
ZHANG Huan-huan;FANG Jie;MAO Shi-ying;YANG Yang;XIE Hui;YU Chen-huan(Key Lab of ExperimentalAnimal and Safety Research of Zhejiang Province,Hangzhou 310013,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2019年第4期514-518,共5页
Chinese Pharmacological Bulletin
基金
浙江省公益技术应用研究项目(No 2016C37134
No.2015C33185)
浙江省医药卫生科研项目(No 2015KYB091)
浙江省实验动物基因工程创新学科建设项目(No 201604)
