紫花牡荆素通过miR-148a-3p/Wnt1信号通路抑制肝癌MHCC97H细胞的迁移和侵袭  

Casticin inhibits migration and invasion of hepatocellular carcinoma MHCC97H cells through miR-148a-3p/Wnt1 signaling pathway

作　　者：娄磊 王靓厚 王智 李翔 LOU Lei;WANG Liang-hou;WANG Zhi;LI Xiang(Dept of Preclinical Medicine,Medical College,Hunan Normal University,Changsha 410013,China)

机构地区：[1]湖南师范大学医学院基础医学系,湖南长沙410013

出　　处：《中国药理学通报》2023年第5期868-875,共8页Chinese Pharmacological Bulletin

基　　金：湖南省自然科学基金面上项目(No 2021JJ30462)。

摘　　要：目的用紫花牡荆素(casticin,CAS)处理MHCC97H细胞,观察其迁移和侵袭能力的变化,并初步探讨CAS的分子作用机制。方法CCK-8试剂盒检测不同浓度CAS对MHCC97H细胞活力的影响;分组开展细胞迁移和侵袭实验,评估MHCC97H细胞的迁移和侵袭能力;逆转录荧光定量PCR(RT-qPCR)检测CAS处理后,MHCC97H细胞的miR-148a-3p和Wnt1 mRNA表达变化;迁移侵袭相关蛋白(MMP2、MMP9)和Wnt1蛋白表达量采用Western blot检测;Dual-Luciferase报告基因检测miR-148a-3p与Wnt13′-UTR的结合情况。结果CAS明显抑制MHCC97H细胞的生存活力,其对这种细胞的IC_(50)约为10.0μmol·L^(-1),亚细胞毒浓度的CAS(3.0μmol·L^(-1))可减弱这种细胞的迁移和侵袭能力;miR-148a-3p mimic或CAS均能抑制MHCC97H细胞迁移和侵袭能力,且两者联合处理后的抑制作用强于单独使用;过表达Wnt1能有效减弱CAS的抑制作用;CAS能明显上调MHCC97H细胞miR-148a-3p表达,同时Wnt1、MMP2和MMP9的表达呈现下降;Dual-Luciferase报告基因发现,miR-148a-3p可以直接靶向Wnt13′-UTR。结论CAS可通过miR-148a-3p/Wnt1信号通路减弱肝癌细胞的迁移和侵袭。Aim To investigate the effect of casticin(CAS)on the migration and invasion of MHCC97H cells and preliminarily explore the molecular mechanism.Methods CCK-8 kit was used to detect the effect of different concentrations of CAS on the viability of MHCC97H cells;cell migration and invasion assays were carried out in groups to assess the migration and invasion ability of MHCC97H cells;reverse transcription fluorescence quantitative PCR(RT-qPCR)was performed to detect the miR-148a-3p and Wnt1 mRNA expression in MHCC97H cells after CAS treatment;migration-invasion related proteins(MMP2,MMP9)and Wnt1 protein expression were detected by Western blot;Dual-Luciferase reporter gene was used to detect the binding of miR-148a-3p to Wnt13′-UTR.Results CAS significantly inhibited the viability of MHCC97H cells.The IC_(50)of CAS for MHCC97H cells was approximately 10.0μmol·L^(-1).Subcytotoxic concentrations of CAS(3.0μmol·L^(-1))attenuated the migration and invasion ability of MHCC97H cells;both miR-148a-3p mimic or CAS inhibited the migration and invasion ability of MHCC97H cells,and the inhibitory effect was stronger after combined treatment than alone;overexpression of Wnt1 effectively attenuated the inhibitory effect of CAS;CAS significantly up-regulated miR-148a-3p expression in MHCC97H cells,while the expression of Wnt1,MMP2 and MMP9 showed a decrease;Dual-Luciferase reporter gene found that miR-148a-3p could directly target Wnt13′-UTR.Conclusion CAS can attenuate the migration and invasion of hepatocellular carcinoma cells through miR-148a-3p/Wnt1 signaling pathway.

关 键 词：紫花牡荆素 肝癌 MHCC97H细胞 miR-148a-3p/Wnt1信号通路 细胞迁移 细胞侵袭 

分 类 号：R282.71[医药卫生—中药学] R394.2[医药卫生—中医学] R73-37R735.7