Rap1型
MAPK/ERK通路
转染
血管平滑肌
细胞生长
细胞生物学
信号转导
鸟嘌呤核苷酸交换因子
细胞迁移
化学
免疫印迹
间质细胞
癌症研究
生物
分子生物学
细胞
细胞培养
生物化学
内分泌学
遗传学
平滑肌
基因
作者
Li Qin,Yunfei Teng,Jian Wang,Miao Yu,Yiqing Li,Hong Zheng
标识
DOI:10.1016/j.prp.2018.04.007
摘要
Rap1 is involved in a multitude of cellular signal transduction pathways, which has extensively been linked to cell proliferation and migration. It has been shown to be important in the regulation of physiological and pathological processes. The present study aims to elucidate its detailed mechanistic in proliferation and migration.Vascular smooth muscle cells (VSMCs) were transfected with pcDNA3.1(empty vector), pcDNA3.1 containing Myc-Tagged-Rap1V12 (Rap1V12) or pcDNA3.1 containing Flag-Tagged-Rap1GAP (Rap1GAP).The cells were presence or absence with 8CPT-2'OMe-cAMP or SDF-1 before transfection. The proliferation and migration were examined by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) and transwell analysis, respectively. Afterwards, western blot was performed to detect the expression of ERK, phosphorylated-ERK, Rap1, Rap1GAP and Rap1GTP.The results showed that proliferation, migration and the expression of Rap1, Rap1GAP, p-EKR were boosted in treatment of Rap1V12-transfection. However, Rap1GAP presented the opposite effects. Subsequently, VSMCs were pretreatment with stimulators Rap1 guanine exchange factor (Rap1GEF), 8CPT-2'OMe-cAMP and stromal cell-derived factor 1 (SDF-1), then transfected with different vectors and the expression of Rap1, Rap1GAP and p-EKR were obviously decreased.Taken together, these findings indicated for the first time that Rap1 was essential for the VSMCs in proliferation and migration by ERK signaling pathway.
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