Curcumin协同ABT-737对肝癌细胞上皮间质转化的抑制作用及相关机制初步研究

肝脏 ›› 2022, Vol. 27 ›› Issue (1): 86-90.

Curcumin协同ABT-737对肝癌细胞上皮间质转化的抑制作用及相关机制初步研究

郑锐年, 孙成晖, 贾筠, 林顺欢, 林钦雄, 刘淳, 郝艳艳, 潘学兵, 何宇, 邵俊伟

523000 广东东莞广东省南方医科大学附属东莞医院肿瘤内科,东莞市肿瘤临床医学研究所(郑锐年,贾筠,林顺欢,林钦雄,刘淳,郝艳艳,潘学兵,何宇,邵俊伟);广东省阳春市人民医院(孙成晖)

收稿日期:2021-02-10 出版日期:2022-01-31 发布日期:2022-02-11
基金资助:
广东省医学科学技术研究基金项目(B2019080)

The combined inhibiting effect and related mechanism of Curcumin and ABT-737 on epithelial-mesenchymal transition of human liver cancer cell

ZHENG Rui-nian¹, SUN Cheng-hui², JIA Yun¹, LIN Shun-huan¹, LIN Qin-xiong¹, LIU Chun¹, HAO Yan-yan¹, PAN Xue-bing¹, HE Yu¹, SHAO Jun-wei¹

1. Dongguan People’s Hospital,Dongguan 523000, Guangdong, China;
2. Yangchun City People's Hospital, Yangchun 529600, Guangdong, China

Received:2021-02-10 Online:2022-01-31 Published:2022-02-11

摘要/Abstract

摘要： 目的探讨Curcumin协同ABT-737对肝癌7404细胞上皮间质转化(epithelial- mesenchymal transition,EMT)的抑制作用及机制研究。方法 2 μmol/L Curcumin、5 μmol/L ABT-737或2 μmol/L Curcumin + 5 μmol/L ABT-737作用肝癌7404细胞后,观察细胞形态的变化,细胞划痕实验检测7404细胞的迁移能力,蛋白质印迹法检测7404细胞中E-cadherin、Vimentin、N-cadherin、ZEB1蛋白的表达和p-beta-catenin、p-JNK、Snail、Twist蛋白的表达。构建肝癌动物模型(Alb-Cre;P53^f/f;Ras),在苏木精伊红染色下观察肝转移瘤情况,并对在肝脏形成的转移灶进行数量统计。结果相比2 μmol/L Curcumin、5 μmol/L ABT-737,2 μmol/L Curcumin + 5 μmol/L ABT-737作用后,7404细胞梭形化明显减少(P<0.05),细胞迁移能力显著降低(P<0.05),Vimentin、N-cadherin、ZEB1蛋白的表达水平明显抑制(P<0.05),E-cadherin蛋白的表达水平明显上调(P<0.05),beta-catenin和JNK的磷酸化水平明显上调(P<0.05),beta-catenin下游靶基因Snail、Twist的表达水平明显抑制(P<0.05)。相比对照组,Curcumin + ABT-737作用后,肝癌动物模型(Alb-Cre;P53^f/f;Ras)中肝转移瘤数量明显减少(P<0.05)。结论 Curcumin协同ABT-737能抑制肝癌细胞上皮间质转化,JNK-beta-catenin可能参与EMT转化的抑制。

关键词: 肝癌, Curcumin, ABT-737, 上皮间质转化

Abstract: Objective To explore the combined inhibiting effect and related mechanism of Curcumin and ABT-737 on epithelial-mesenchymal transition (EMT) of human liver cancer cell.Methods After treatment with 2 μmol/L Curcumin, 5 μmol/L ABT-737 alone or 2 μmol/L Curcumin in combination with 5 μmol/L ABT-737, the morphological change of 7404 cells was observed under an inverted microscope, the cell migration was analyzed by Wound Healing, the expression levels of Vimentin, N-cadherin, ZEB1 and E-cadherin proteins in 7404 cells were detected by Western blotting. The expression levels of p-beta-catenin, p-JNK, Snail and Twist proteins in 7404 cells were also detected by Western blotting. The animal models of liver cancer(Alb-Cre; P53^f/f; Ras)was established. The numbers of metastatic tumor in liver was observed and calculated under hematoxylin eosin stain.Results As compared with the 7404 cells with treatment of 2 μmol/L Curcumin or 5 μmol/L ABT-737, the numbers of 7404 cells that changed shape to spindle-like after treatment with 2 μmol/L Curcumin + 5 μmol/L ABT-737 were obviously reduced (P<0.05), with significantly decline of cell migration ability. Compared with the 2 μmol/L Curcumin or 5 μmol/L ABT-737 group, the expression levels of E-cadherin protein were significantly increased while the expression levels of Vimentin, N-cadherin and ZEB1 protein were significantly decreased in 7404 cells after treatment with 2 μmol/L Curcumin and 5 μmol/L ABT-737 (P<0.05). What’s more, as compared with the 7404 cells treated with the 2 μmol/L Curcumin or 5 μmol/L ABT-737 group,the expression levels of p-beta-catenin and p-JNK protein were significantly increased while the expression levels of Snail and Twist protein were decreased significantly in the 7404 cells after treatment with 2 μmol/L Curcumin + 5 μmol/L ABT-737 (P<0.05). The numbers of metastatic tumor in liver were decreased significantly in the animal models of liver cancer (Alb-Cre; P53^f/f; Ras) after Curcumin and ABT-737 treatment than those in the animal models without any treatment (as a control) (P<0.05).Conclusion The combination treatment of curcumin and ABT-737 may inhibit the epithelial-mesenchymal transition of human liver cancer cell, and JNK-beta-catenin signaling pathway may be involved in EMT.

Key words: Liver cancer, Curcumin, ABT-737, Epithelial-mesenchymal transition

郑锐年, 孙成晖, 贾筠, 林顺欢, 林钦雄, 刘淳, 郝艳艳, 潘学兵, 何宇, 邵俊伟. Curcumin协同ABT-737对肝癌细胞上皮间质转化的抑制作用及相关机制初步研究[J]. 肝脏, 2022, 27(1): 86-90.

ZHENG Rui-nian, SUN Cheng-hui, JIA Yun, LIN Shun-huan, LIN Qin-xiong, LIU Chun, HAO Yan-yan, PAN Xue-bing, HE Yu, SHAO Jun-wei. The combined inhibiting effect and related mechanism of Curcumin and ABT-737 on epithelial-mesenchymal transition of human liver cancer cell[J]. Chinese Hepatolgy, 2022, 27(1): 86-90.

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