摘要
[目的]探讨萝卜硫素与索拉菲尼联合对肝癌HepG2细胞生长抑制及凋亡的作用,并观察其可能的机制。[方法]细胞计数试剂盒8(CCK8)法分析了萝卜硫素、索拉菲尼及两者联合给药对HepG2细胞增殖抑制率的影响,计算每种药物的半数抑制浓度(IC50)及联合时的联合指数(CI);流式细胞仪检测了HepG2细胞凋亡情况;免疫蛋白印记(Western Blot)检测了细胞周期蛋白D1(ClinD1)、C-myc、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、磷酸化核因子-κB(p-NF-κB)及磷酸化核因子κB抑制蛋白α(p-IκBα)蛋白表达水平。[结果]萝卜硫素与索拉菲尼联合对HepG2细胞增殖抑制作用和凋亡诱导作用显著高于各单独给药(P<0.05),还表现出浓度依赖性,呈现出明显的协同作用(CI值<1)。Western Blot结果显示,相比于对照组及单独给药组,联合给药能显著抑制原癌基因[细胞周期蛋白D1(ClinD1)和C-myc]、抗凋亡蛋白Bcl-2、p-NF-κB及磷酸化IκBα蛋白的表达,诱导促凋亡蛋白Bax的表达(P<0.05)。[结论]萝卜硫素与索拉菲尼联合对HepG2细胞具有协同抗肿瘤作用,机制可能与诱导细胞凋亡及抑制NF-κB信号通路活化有关。
[Objective]To observe the effects of sulforaphane combined with sorafenib on the hepatocellular carcinoma HepG2 cells and study its mechanism.[Methods]The anti-proliferation effects of either sulforaphane or sorafenib alone or their combination on HepG2 cells were assessed by CCK8 assay.The IC50 values of two single drug and the CI values were calculated.The apoptosis ratios were analyzed by flow cytometer.The expression of ClinD1,C-myc,Bcl-2,Bax,p-NF-κB and p-IκBαwere detected by Western blotting.[Results]The anti-proliferation and apoptosis induction effects were more remarkably than the two single drugs when combination with sulforaphane and sorafenib(P<0.05).It appeared in a concentration-dependent manner.The CI values of sulforaphane combinated with sorafenib were all less than alone,which had a synergistic effect.Western blotting showed that sulforaphane combinated with sorafenib could significantly decrease the expression levels of ClinD1,C-myc,Bcl-2,p-NF-κB and p-IκBαand increase the expression level of Bax in HepG2 cells,as compared with the control and the single drugs(P<0.05).[Conclusion]The combination of sulforaphane and sorafenib performs a synergistic anti-tumor effect on HepG2 cells,and its mechanism underlying may be related to apoptosis and down-regulation the activation of NF-κB pathway.
作者
黄建峰
魏小栋
翟东升
HUANG Jianfeng;WEI Xiaodong;ZHAI Dongsheng(Department of Hepato-Pancreato-Biliary Surgery,Affiliated Hospital of Hubei University for Nationalities,Enshi 445000,China;Department of Cardiothoracic Surgery,Affiliated Hospital of Hubei University for Nationalities,Enshi 445000,China)
出处
《天津中医药》
CAS
2019年第12期1222-1226,共5页
Tianjin Journal of Traditional Chinese Medicine
基金
湖北省自然科学基金(ZRMS-2018001241)。
关键词
萝卜硫素
索拉菲尼
肝癌细胞
协同作用
sulforaphane
sorafenib
hepatocellular carcinoma cells
synergistic effect
