健脾化瘀方调控circβ-catenin/Wnt通路介导肝癌细胞上皮间质转化的机制  

Mechanism of Jianpi Huayu Formula regulating epithelial mesenchymal transition of hepatocellular carcinoma cell via circβ-catenin/Wnt pathway

作　　者：方崇锴 王冀南 刘慧琳 刘丽华 冯坤良 章颖 王俊岩 林宏通 钟崇[2] FANG Chong-kai;WANG Ji-nan;LIU Hui-lin;LIU Li-hua;FENG Kun-liang;ZHANG Ying;WANG Jun-yan;LIN Hong-tong;ZHONG Chong(Guangzhou University of Chinese Medicine,Guangzhou 510405,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Guangdong Province Traditional Chinese Medical Hospital,Guangzhou 510120,China)

机构地区：[1]广州中医药大学,广州510405 [2]广州中医药大学第一附属医院,广州510405 [3]广东省中医院,广州510120

出　　处：《中华中医药杂志》2022年第5期2513-2518,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81873303),广东省教育厅普通高校重点项目(No.2019 KZDXM045),广东省自然科学基金面上项目(No.2019A1515011013)。

摘　　要：目的:探讨健脾化瘀方抑制肝癌细胞增殖和侵袭的作用及机制。方法:制备健脾化瘀方冻干粉,干预人肝癌HepG2细胞后,CCK-8试剂检测细胞活力,划痕实验检测细胞的迁移能力,Transwell实验检测细胞的侵袭能力。qPCR检测circβ-catenin mRNA表达水平的变化,Western Blot检测细胞上皮间质转化(EMT)相关蛋白的表达。免疫荧光检测细胞β-catenin、N-cadherin的表达情况。结果:健脾化瘀方抑制肝癌HepG2细胞的增殖,随着药物浓度的升高,细胞活力显著降低。与空载质粒(p-vector)组比较,转染circβ-catenin质粒后,细胞迁移能力和侵袭能力显著提高(P<0.01),circβ-catenin mRNA表达水平显著升高(P<0.01),Wnt2、β-catenin、Vimentin、N-cadherin蛋白水平显著升高(P<0.05,P<0.01),E-cadherin蛋白表达水平显著降低(P<0.05)。与p-vector组或circβ-catenin-OE组比较,健脾化瘀方干预后,抑制肝癌细胞发生EMT,下调Wnt2、β-catenin、Vimentin、N-cadherin蛋白水平(P<0.05,P<0.01),上调E-cadherin蛋白水平(P<0.01)。结论:健脾化瘀方能够抑制HepG2细胞的增殖及侵袭,其机制可能是通过调节circβ-catenin/Wnt通路介导EMT发挥作用。Objective:To investigate the effects and mechanism of Jianpi Huayu Formula on inhibiting the proliferation and invasion of hepatocellular carcinoma cell.Methods:The freeze-dried powder of Jianpi Huayu Formula was prepared.After intervention with human HepG2 cells of liver cancer,the CCK-8 reagent detected the cell viability,the scratch test detected the cell migration ability,and the Transwell test detected the cell invasion ability.qPCR was used to detect changes in the mRNA expression level of circβ-catenin,and Western Blot was used to detect the expression of EMT-related proteins in cells.Immunofluorescence was used to detect the expression ofβ-catenin and N-cadherin in cells.Results:Jianpi Huayu Formula inhibited the proliferation of HepG2 cells of liver cancer.With the increase of drug concentration,the cell viability decreased significantly.Compared with the p-vector group,after transfection with circβ-catenin plasmid,the cell migration and invasion ability was significantly improved(P<0.01),the mRNA expression level of circβ-catenin was greatly increased(P<0.01).The protein levels of Wnt2,β-catenin,Vimentin and N-cadherin increased significantly(P<0.05,P<0.01),and the protein expression levels of E-cadherin decreased significantly(P<0.05).Compared with the p-vector group or circβ-catenin-OE group,Jianpi Huayu Formula inhibited the occurence of EMT in liver cancer cells,down-regulated the proteins levels of Wnt2,β-catenin,Vimentin,N-cadherin(P<0.05,P<0.01),upregulated the protein level of E-cadherin(P<0.01).Conclusion:Jianpi Huayu Formula can inhibit the proliferation and invasion of HepG2 cells,possibly by regulating EMT mediated by circβ-catenin/Wnt pathway.

关 键 词：健脾化瘀方 肝癌 circβ-catenin WNT通路 上皮间质转化 机制 迁移 侵袭 

分 类 号：R285[医药卫生—中药学]