姜黄素通过Wnt2/β-catenin通路逆转食管癌Eca-109/VCR细胞多药耐药性研究  

Curcumin reversed multi-drug resistance of esophageal carcinoma in Eca-109/VCR cell line through Wnt2/β-catenin pathway

作　　者：任海玉 孙剑经[2] 李多[2] 牛树荣 刘华 罗强 张林西 REN Hai-yu;SUN Jian-jing;LI Duo;NIU Shu-rong;LIU Hua;LUO Qiang;ZHANG Lin-xi(Postgraduate School of Hebei North University,Hebei North University,Zhangjiakou 075000,China;Digestion Dept,1st Affiliated Hospital,Hebei North University,Zhangjiakou 075000,China;Life Science Research Centre of Hebei North University,Zhangjiakou 075000,China)

机构地区：[1]河北北方学院研究生学院,河北张家口075000 [2]河北北方学院附属第一医院消化内科,河北张家口075000 [3]河北北方学院生命科学研究中心,河北张家口075000

出　　处：《中国药理学通报》2018年第10期1455-1460,共6页Chinese Pharmacological Bulletin

基　　金：河北省自然科学基金资助项目(No H2014405033);河北北方学院自然科学课题项目(No ZD201414)

摘　　要：目的研究姜黄素(curcumin,Cur)对人食管癌耐药细胞株Eca-109/VCR多药耐药的逆转作用,并探讨其可能的逆转机制。方法 CCK-8法检测不同浓度Cur和VCR对Eca-109/VCR细胞增殖的影响,以及Cur(20μmol·L^(-1))对Eca-109/VCR细胞多药耐药的逆转作用;FCM检测细胞凋亡率;ELISA法检测P-gp蛋白的表达水平;实时荧光定量PCR分析Wnt2、β-catenin mRNA的相对表达量;Western blot检测Wnt2、β-catenin、MMP-2、HMGB1的蛋白表达水平。结果随着Cur浓度的增加,细胞抑制率逐渐升高,呈剂量依赖性。Cur(20μmol·L^(-1))与不同浓度VCR联合作用可明显提高细胞增殖抑制率,逆转倍数为3.49。Cur(20μmol·L^(-1))与VCR(2.0 mg·L^(-1))联合作用可明显提高细胞凋亡率,降低P-gp蛋白含量,下调Wnt2、β-catenin mRNA的相对表达,并抑制Wnt2、β-catenin、MMP-2、HMGB1蛋白的表达。结论 Cur具有逆转食管癌细胞多药耐药的作用,其作用机制可能与下调Wnt2、β-catenin的表达,抑制Wnt2/β-catenin通路活性,以及下调侵袭相关蛋白MMP-2、HMGB1的表达有关。To study the reversal effect of curcumin(Cur)on vincristine(VCR)-resistance in human esophageal carcinoma Eca-109/VCR cells as well as its mechanism.Methods The esophageal carcinoma drug-resistant Eca-109/VCR cells were treated with different concentrations of Cur and VCR.The proliferation of Eca-109/VCR cells were detected by CCK-8 assay,and the concentration of Cur screened by CCK-8 assay.The apoptosis rate was detected by flow cytometry(FCM).The level of P-glycoprotein(P-gp)was measured by enzyme-linked-immunosorbent assay(ELISA).The level of Wnt2 andβ-catenin mRNA were detected by quantitative real-time polymerase chain reaction.The relative protein expressions ofWnt2,β-catenin,MMP-2 and HMGB1 were analyzed by Western blot.Results The proliferation inhibitory rates were positively correlated with the concentration of curcumin,in a dose-dependent manner.The combination of Cur(20μmol·L-1)and different concentrations of VCR could significantly increase the proliferation inhibitory rates of cells.The drug-resistant reversal fold was 3.49.The combination of Cur(20μmol·L-1)and VCR(2.0 mg·L-1)could markedly increase the apoptosis ratios of cells,reduce the protein expression of P-gp,down-regulate the level of Wnt2 andβ-catenin mRNA,and decrease the relative protein level of Wnt2,β-catenin,MMP-2 and HMGB1.Conclusions Cur can reverse multi-drug resistance of Eca-109/VCR cells.The mechanism may be related to the down-regulation of Wnt2 andβ-catenin,which can inhibit the activity of Wnt2/β-catenin pathway.The down-regulation of MMP-2 and HMGB1 may also be one of the possible mechanisms.

关 键 词：食管癌 Wnt2/β-catenin通路 姜黄素 长春新碱 多药耐药 实时荧光定量PCR 

分 类 号：R282.71[医药卫生—中药学] R329.24[医药卫生—中医学]