中药化学成分库中结肠癌BRAF^(V600E)抑制剂的筛选  

Screening of monomers from traditional Chinese medicine for specifically inhibiting BRAF^(V600E) CT26 cells

作　　者：张欢欢[1] 方杰[1] 毛诗莹 杨扬 谢珲[1] 余陈欢[1] 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)

机构地区：[1]浙江省医学科学院浙江省实验动物中心浙江省实验动物与安全性研究重点实验室,浙江杭州310013

出　　处：《中国药理学通报》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)

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.

关 键 词：BRAF^V600E CT26细胞系 细胞增殖 MEK/ERK信号通路 中药单体化合物 抑制剂 

分 类 号：R-332[医药卫生] R282.71