槲皮素通过G3BP1调控Wnt信号通路抑制宫颈癌细胞增殖  

Quercetin Inhibits Proliferation of Cervical Cancer Cells by Regulating Wnt Signaling Pathway through G3BP1

作　　者：向秋玲[1] 李雪兰[1] 黄薇霖[1] 杨杰[1] Xiang Qiuling;Li Xuelan;Huang Weilin;Yang Jie(Department of Medicine,Changde Vocational and Technical College,Changde 415003)

机构地区：[1]常德职业技术学院医学系,常德415003

出　　处：《中药药理与临床》2020年第4期102-108,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：湖南省教育厅科学研究项目(编号:17C0205);常德市科技事业发展基金项目(编号:2016KZ32);常德职业技术学院院级科研项目(编号:ZY1607)。

摘　　要：目的:探讨槲皮素通过G3BP1调控Wnt/β-Catenin通路抑制宫颈癌细胞增殖的作用机制。方法:建立SiHa细胞裸鼠皮下移植瘤模型,设置模型对照组、槲皮素25 mg/kg、50 mg/kg、100 mg/kg组,检测给药第7 d、14 d、21 d瘤体积变化,给药结束后取肿瘤组织称重,Western blot检测瘤组织中GTP酶激活蛋白-Src同源结构域3-结合蛋白1(G3BP1)、β-连环蛋白(β-Catenin)水平。采用CCK-8法分析5μmol/L^320μmol/L槲皮素作用48 h对SiHa细胞增殖的抑制作用,并计算半数抑制浓度(IC50);将25、50及100μmol/L槲皮素作用于SiHa细胞,绘制7 d生长曲线;观察细胞集落形成情况;采用Western blot法检测细胞G3BP1、β-Catenin、细胞周期蛋白D1(cyclinD1)、Myc原癌基因(c-Myc)蛋白表达水平;敲低或上调G3BP1的表达,观察其对抑制率以及G3BP1、β-Catenin、cyclinD1、c-Myc蛋白水平的影响。结果:槲皮素25 mg/kg、50 mg/kg、100 mg/kg组给药第14 d、21 d瘤体积明显小于模型对照组,与模型对照组比较,槲皮素50、100 mg/kg组能明显降低瘤体质量(P<0.05)。25、50、100 mg/kg的槲皮素能明显下调瘤体G3BP1、β-Catenin蛋白的表达(P<0.05或P<0.01)。CCK-8结果显示,槲皮素对宫颈癌SiHa细胞的增殖均具有抑制作用,并伴随槲皮素浓度的增加,抑制作用明显增加,IC50为(84.19±5.12)μmol/L;生长曲线提示槲皮素25、50、100μmol/L对SiHa细胞生长具有明显的抑制作用;显著抑制SiHa细胞的集落形成;与模型对照组比较,槲皮素25、50、100μmol/L可下调G3BP1、β-Catenin、cyclinD1、c-Myc蛋白表达;敲低G3BP1的表达,槲皮素50μmol/L显著增加抑制率,下调β-Catenin、cyclinD1、c-Myc蛋白表达,上调G3BP1的表达,槲皮素50μmol/L显著降低抑制率,上调β-Catenin、cyclinD1、c-Myc蛋白表达(P<0.05或P<0.01)。结论:槲皮素能抑制宫颈癌细胞增殖,其机制可能是通过靶向G3BP1调控Wnt/β-Catenin通路有关。Objective: To investigate the mechanism of quercetin on regulating Wnt/β-Catenin pathway through G3 BP1 to inhibit the growth of cervical cancer cells. Methods: SiHa nude mouse subcutaneously transplanted tumor model was established,mice were randomly divided into the model group,25 mg/kg,50 mg/kg and 100 mg/kg quercetin groups.The tumor volume changes in each group on the 7 th, 14 d, and 21 d of drug administration were measured.After administration,tumor tissues were collected and weighed,the levels of G3 BP1 and β-Catenin phosphorylation were detected in the tumor tissues by Western blot. The inhibitory effects of 5 μmol/L^320 μmol/L quercetin on the proliferation of SiHa cells were analyzed by CCK-8 method, and the half-inhibitory concentration(IC50) was calculated. SiHa cells was treated with quercetin(25, 50 and 100μmol/L) for 7 d to draw the growth curve. The colony formation was observed,the expression levels of G3 BP1, β-Catenin, cyclinD1 and c-Myc protein were detected by Western blot. After knockdown or up-regulation of G3 BP1 expression,the inhibition rate and levels of G3 BP1,β-Catenin, cyclinD1, and c-Myc protein were observed. Results: Compared with the model group, the tumor volumes in quercetin groups(25 mg/kg,50 mg/kg,100 mg/kg) on the 14 th and 21 d of drug administration were significantly decreased, the tumor masses were significantly decreased in quercetin groups(50, 100 mg/kg)(P<0.05), the expression levels of G3 BP1 and β-Catenin were down regulated by quercetin( 25, 50, and 100 mg/kg). CCK-8 results showed all doses of quercetin inhibited the growth of cervical cancer SiHa cells,in a concentration dependent manner, and the IC50 was 84.19±5.12 μmol/L. The growth curve indicated that quercetin( 25, 50, 100 μmol/L) significantly inhibited the growth of SiHa cells and the colony formation of SiHa cells. Compared with the model group,quercetin(25, 50, 100 μmol/L) down-regulated protein levels of G3 BP1, β-Catenin, cyclinD1 and c-Myc. After the knockdown of G3 BP1 expression, 50 μ

关 键 词：槲皮素 GTP酶激活蛋白-Src同源结构域3-结合蛋白1 Wnt/β-连环蛋白通路 宫颈癌 抑制率 

分 类 号：R285.5[医药卫生—中药学]