黄芩苷对肝癌细胞GJIC及Cx26、Cx43表达的影响  

Treatment with baicalin up-regulates the expression of connexion 26 and connexion 43 in human hepatocellular carcinoma cell line SMMC-7721

作　　者：郭昱[1] 王娜[2] 高咏梅[3] 姚金锋[1] 李元[1] 尹崇娇[1] 张文娟[1] 

机构地区：[1]河北医科大学第二医院消化内科,河北省石家庄市050000 [2]保定市第三医院消化内科,河北省保定市071000 [3]河北北方学院附属第一医院消化内科,河北省张家口市075000

出　　处：《世界华人消化杂志》2012年第33期3197-3202,共6页World Chinese Journal of Digestology

基　　金：河北省自然科学基金资助项目;No.C2009001123~~

摘　　要：目的:探讨黄芩苷对肝癌细胞缝隙连接细胞间通讯(gap junction intercellular communication,GJIC)及细胞间隙连接蛋白26(connexion26,Cx26)、细胞间隙连接蛋白43(connexion43,Cx43)表达的影响.方法:人肝癌细胞株SMMC-7721分为黄芩苷10、20、40mg/L组和对照组.划痕染料示踪技术检测GJIC的变化.用RT-PCR法检测肝癌细胞Cx26、Cx43基因表达,用Westernblot法检测Cx26蛋白表达,用免疫细胞化学检测Cx43蛋白表达.结果:对照组细胞的染料局限于划痕两侧的细胞内,无明显的荧光染料传输现象.随着黄芩苷浓度的增强,LY染料传输范围逐渐增大.黄芩苷10、20、40mg/L各浓度组Cx26mRNA表达水平与对照组比较显著提高(0.148±0.111,10.253±0.222,17.283±0.024vs0.138±0.111;P<0.05).Cx26蛋白表达量与对照组比较明显增加(0.516±0.029,0.759±0.020,1.019±0.076vs0.367±0.029;P<0.05).黄芩苷各浓度组Cx43mRNA表达量与对照组比较无显著变化.而Cx43蛋白的表达水平与对照组比较明显增加(5.512±0.003,5.844±0.046,6.216±0.015vs4.316±0.032;P<0.05).结论:黄芩苷促进肝癌细胞Cx26及Cx43表达,导致肝癌细胞GJIC功能的恢复,这很可能是黄芩苷抑制肿瘤生长的分子机制之一.AIM： To investigate the effect of treatment with baicalin on the expression of connexion 26 （Cx26） and connexion 43 （Cx43） in human hepatocellu- lar cell line SMMC-7721. METHODS： SMMC-7721 cells were randomly divided into 4 groups： normal control group, low-dose （10 mg/L） baicalin group, medium- dose （20 rag/L） baicalin group, and high-dose （40 rag/L） baicalin group. Gap junction intercellularcommunication （GJIC） was measured by scrap loading/dye transfer assay （SL/DT）. The expres- sion of Cx26 and Cx43 mRNAs in SMMC-7721 cells was determined by RT-PCR. The expres- sion of Cx26 protein was detected by Western blot, and that of Cx43 protein was detected by immunohistochemistry. RESULTS： Compared to the normal control group, the expression of Cx26 mRNA and pro- tein was significantly enhanced in SMMC-7721 cells treated with low, medium and high con- centrations of baicalin （mRNA： 0.148 ＋ 0.111, 10.253 ＋ 0.222, 17.283 ＋ 0.024 vs 0.138 ＋ 0.111; all P 〈 0.05; protein： 0.516 ＋ 0.029, 0.759 ＋ 0.020, 1.019 ＋ 0.076 vs 0.367 ＋ 0.029; all P 〈 0.05）. Compared to the normal control group, the expression of Cx43 mRNA showed no significant changes, but the expression of Cx43 protein was significantly enhanced in SMMC-7721 cells treated with dif- ferent concentrations of baicalin. CONCLUSION： Restoration or enhancement of GJIC induced by up-regulation of Cx26 and Cx43 is likely to be an important molecular mechanism by which baicalin inhibits tumor growth.

关 键 词：黄芩苷 肝癌 肝癌细胞缝隙连接细胞间通讯 细胞间隙连接蛋白26 细胞间隙连接蛋白43 

分 类 号：R285[医药卫生—中药学] R735.7[医药卫生—中医学]