中国药理学与毒理学杂志2025，Vol.39Issue(4)：252-259,8.DOI:10.3867/j.issn.1000-3002.2025.04.002
α吡喃酮类化合物Rasfonin对鸟苷酸交换因子SOS1表达的调控作用及其潜在机制
Effects of Rasfonin,an α-pyrone derivative,on SOS1 expression and underlying mechanisms
摘要
Abstract
OBJECTIVE To investigate the regulatory effect of Rasfonin on SOS1(Son of Seven-less,one of the major guanylate exchange factors)expressions and the underlying mechanism.METHODS① Human cancer cells MCF-7(breast cancer cells,KRASWT wild-type),Calu-1(non-small cell lung cancer,KRASG12C mutation),and UM-UC-3(bladder metastatic cell carcinoma,KRASG12C mutation)were divided into the control group and Rasfonin(1,5,10 and 15 μmol·L-1)treated groups.CCK-8 assay was used to observe the effects of Rasfonin on the proliferation of MCF-7,Calu-1,and UM-UC-3 cells after 24 h of Rasfonin treatment.In addition,these cells were divided into the control group,EGF stimulation group(EGF 50 μg·L-1,stimulated for 5 min),and Rasfonin treated groups(pretreated with 5 and 10 μmol·L-1 Rasfonin before 5 min EGF stimulation).Quantitative real-time PCR(real-time fluores-cence PCR)and Western blotting were employed to identify the expression levels of SOS1 mRNA and protein in MCF-7,Calu-1 and UM-UC-3 cells.② The co-expression systems of KRAS and SOS1 were established by transfecting plasmids(KRAS-NC,KRASWT,KRASG12C and SOS1)into 293T cells that were divided into the control group and Rasfonin(1,5 and 10 μmol·L-1)treated group.The dual luciferase reporter gene assay was used to evaluate the effects of Rasfonin on activities of the SOS1 promoter.Moreover,293T cells were divided into the EGF stimulation group(EGF 50 μg·L-1,stimulated for 5 min)and Rasfonin treated groups(12 h of treatment with 10 μmol·L-1 Rasfonin before 5 min EGF stimula-tion).Western blotting was performed to determine the role of KRASG12C protein in the inhibition of Rasfonin on SOS1 expression.RESULTS ① Compared with the control group,Rasfonin inhibited the prolifera-tion of Calu-1 and UM-UC-3 cells at concentrations of 5,10 and 15 μmol·L-1(IC50 was 8.22 and 4.94 μmol·L-1).But for MCF-7 cells,only 15 μmol·L-1 Rasfonin could decrease their viability(IC50 was 45.15 μmol·L-1).Compared with the EGF stimulation group,mRNA expressions of SOS1 were increased after Rasfonin treatment of 1 h.mRNA expressions of SOS1 were decreased in Calu-1 cells after 3 h of Rasfonin treatment.These changes also occurred after Rasfonin treatment of 3 h and 6 h in UM-UC-3 cells.Further-more,Rasfonin treatment did not influence SOS1 protein expressions in MCF-7 cells,but can signifi-cantly inhibit SOS1 expression of in UM-UC-3 and Calu-1 cells.② Rasfonin had no significant effects on the activity of SOS1 promoter and its protein level in 293T cells when only SOS1 was expressed,but significantly inhibited its activity and its protein level when SOS1 was co-expressed with KRAS protein.CONCLUSION One of the anti-tumor mechanisms of Rasfonin is to inhibit the activity of SOS1 promoter to decrease mRNA and protein expressions of SOS1 through KRASG12C protein.关键词
Rasfonin/SOS1/KRAS突变/肿瘤Key words
Rasfonin/SOS1/KRAS mutation/cancer分类
医药卫生引用本文复制引用
李静,张凯迪,肖智勇,周文霞..α吡喃酮类化合物Rasfonin对鸟苷酸交换因子SOS1表达的调控作用及其潜在机制[J].中国药理学与毒理学杂志,2025,39(4):252-259,8.基金项目
国家自然科学基金(81803431) National Natural Science Foundation of China(81803431) （81803431）
