草苁蓉环烯醚萜苷联合5-氟尿嘧啶对人肝癌HepG2、SK-Hep1细胞上皮间质转化的抑制效果研究  

Inhibitory effects of iridoid glycosides from Boschniakia rossica combined with 5-fluorouracil on human hepatoma HepG2 and SK-Hep1 cells epithelial-mesenchymal transition

作　　者：董学花 延光海[2] 朱洁波 刘莉园 全吉淑[2] 李良昌[2] 尹学哲[1] DONG Xue-hua;YAN Guang-hai;ZHU Jie-bo;LIU Li-yuan;QUAN Ji-shu;LI Liang-chang;YIN Xue-zhe(Affiliated Hospital of Yanbian University,Yanji 133000,China;Yanbian University Medical College,Yanji 133000,China)

机构地区：[1]延边大学附属医院,吉林延吉133000 [2]延边大学医学院,吉林延吉133000

出　　处：《中草药》2020年第17期4498-4505,共8页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81760659)。

摘　　要：目的研究草苁蓉环烯醚萜苷(IGBR)联合5-氟尿嘧啶(5-Fu)抑制转化生长因子-β1(TGF-β1)诱导人肝癌HepG2、SK-Hep1细胞上皮间质转化(Epithelial-mesenchymal transition,EMT),比较其药物的作用疗效。方法采用噻唑蓝(MTT)法检测HepG2、SK-Hep1细胞存活率,计算联合指数(Q值),判断联合用药的相互作用;建立HepG2、SK-Hep1细胞EMT模型,检测细胞黏附率,Westernblotting检测相关的蛋白表达,免疫荧光法检测钙黏附蛋白-E(E-cadherin)和波形纤维蛋白(Vimentin)的表达。结果人肝癌细胞在给药48 h细胞存活率降低(P<0.05),IGBR和5-Fu两者有相加或协同作用;与模型组比较,给药组(IGBR组、5-Fu组、联合组)抑制细胞黏附率(P<0.05);与对照组比较,模型组基质金属蛋白酶2、7、9(MMP2、MMP7、MMP9)、锌指转录因子1、2(Snail、Slug)蛋白表达上调(P<0.05);与模型组比较,给药组(IGBR组、5-Fu组、联合组)MMP2、MMP7、MMP9、Snail、Slug蛋白表达降低(P<0.05);与对照组比较,模型组E-cadherin荧光强度减弱,Vimentin荧光强度增强;与模型组比,给药组(IGBR组、5-Fu组、联合组)E-cadherin荧光强度增强,Vimentin荧光强度减弱。结论 IGBR和5-Fu可以抑制肝癌EMT,联合用药对HepG2细胞作用相加;对SK-Hep1细胞具有协同效果,SK-Hep1细胞用药疗效优于HepG2细胞。Objective To study the inhibitory effect of iridoid glycosides from Boschniakia rossica(IGBR) combined with 5-Fu on epithelial-mesenchymal transition induced by TGF-β1 in human hepatoma SK-Hep1 and HepG2 cells, and compare the efficacy of drugs. Methods The survival ability of HepG2 and SK-Hep1 cells was detected by MTT and the combination index(Q value) was calculated to judge the interaction of combined drugs. The EMT model of HepG2 and SK-Hep1 cells was established. The cell adhesion rate was detected by MTT. The expression of matrix metalloproteinase(MMP) MMP2, MMP7, MMP9, Snail, and Slug was detected by Western blotting. The localization and expression intensity of E-cadherin and Vimentin was detected by immunofluorescence. Results MTT showed that compared with the control group, the 5-FU group, IGBR group and combination group cell survival ability were decreased(P < 0.05) at 48 h after administration;IGBR and 5-Fu had an additive or synergistic effect. Compared with the model group, the adhesion rate of 5-FU group, IGBR group and combination group was reduced(P < 0.05). Western blotting results showed that compared with the control group, the expression of MMP2, MMP7, MMP9, Snail, Slug were up-regulated(P < 0.05) in the model group. Compared with the model group, the expression of MMP2, MMP7, MMP9, Snail and Slug were down-regulated(P < 0.05) in 5-FU group, IGBR group and combination group. Compared with the control group, immunofluorescence showed that the E-cadherin fluorescence intensity was decreased in the model group, but the Vimentin fluorescence intensity was increased. Compared with the model group, the E-cadherin fluorescence intensity was increased in 5-FU group, IGBR group and combined group, but the Vimentin fluorescence intensity was decreased. Conclusion IGBR and 5-Fu can inhibit human hepatoma EMT. The combined drugs have the combined effect on HepG2 cells and synergistic effect on SK-Hep1 cells. The therapeutic effect on SK-Hep1 cells is better than HepG2 cells.

关 键 词：草苁蓉 环烯醚萜苷 5-氟尿嘧啶 肝癌 HepG2细胞 SK-Hep1细胞 上皮间质转化 转化生长因子-β1 细胞存活率 细胞黏附率 Western blotting 免疫荧光法 钙黏附蛋白-E 波形纤维蛋白 基质金属蛋白酶 锌指转录因子 协同 

分 类 号：R285.5[医药卫生—中药学]