化瘀散水煎剂通过调控miR-129-5p/FOXC1轴抑制肝癌裸鼠移植瘤生长  

Huayusan decoction inhibited the growth of transplanted liver cancer in nude mice by regulating miR-129-5p/FOXC1 axis

作　　者：许丹[1] 郑超[2] 余首德[1] 严红梅[1] 黄超群 XU Dan;ZHENG Chao;YU Shoude;YAN Hongmei;HUANG Chaoqun(Department of Spleen and Stomach Diseases,Wuhan Hospital of Traditional Chinese Medicine,Hubei Wuhan 430014,China;Department of Surgery,Wuhan Hospital of Traditional Chinese Medicine,Hubei Wuhan 430014,China;Department of Spleen and Stomach Diseases,Hubei Provincial Hospital of Traditional Chinese Medicine,Hubei Wuhan 430070,China;Department of Spleen and Stomach Diseases,Affiliated Hospital of Hubei University of Traditional Chinese Medicine,Hubei Wuhan 430000,China;Institute of Epidemic Diseases,Hubei University of Traditional Chinese Medicine,Hubei Wuhan 430061,China.)

机构地区：[1]武汉市中医医院脾胃病科,湖北武汉430014 [2]武汉市中医医院外科,湖北武汉430014 [3]湖北省中医院脾胃病科,湖北武汉430070 [4]湖北中医药大学附属医院脾胃病科,湖北武汉430000 [5]湖北中医药大学疫病研究所,湖北武汉430061

出　　处：《现代肿瘤医学》2024年第6期1003-1009,共7页Journal of Modern Oncology

基　　金：湖北省武汉市医学科研项目(编号:WZ20C02);国家中医药管理局资助项目(编号:国中医药人教函〔2018〕134号);湖北省武汉市科学技术局曙光计划项目(编号:2022020801020584)。

摘　　要：目的:探讨化瘀散水煎剂通过调控微小RNA-129-5p/叉头框蛋白C1(FOXC1)轴对肝癌裸鼠移植瘤生长的影响。方法:建立肝癌裸鼠移植瘤模型,并分为裸鼠NC组、化瘀散水煎剂组、化瘀散水煎剂+inhibitor-NC组、化瘀散水煎剂+miR-129-5p inhibitor组,每组8只。qRT-PCR法检测miR-129-5p、FOXC1 mRNA表达水平,双荧光素酶报告基因实验验证miR-129-5p与FOXC1靶向调控关系,观察记录裸鼠的瘤重及移植瘤体积,HE染色观察移植瘤组织形态,TUNEL检测肿瘤组织细胞凋亡,免疫组织化学染色检测瘤组织Ki-67、Cleaved Caspase-3、FOXC1表达,Western blot检测Ki-67、Cleaved Caspase-3、FOXC1蛋白表达。结果:miR-129-5p在肝癌细胞中表达显著下降,FOXC1 mRNA表达显著升高,其中HepG2细胞变化最显著(P<0.05);双荧光素酶报告基因实验表明miR-129-5p与FOXC1存在靶向调控关系;与NC组相比,化瘀散水煎剂组肿瘤细胞出现坏死,凋亡数增加,miR-129-5p、Cleaved Caspase-3显著升高,移植瘤质量和体积、FOXC1、Ki-67表达显著降低(P<0.05);下调miR-129-5p逆转化瘀散水煎剂对肝癌移植瘤的抑制作用(P<0.05)。结论:化瘀散水煎剂可能通过上调miR-129-5p从而下调FOXC1表达,进而抑制肝癌移植瘤生长。Objective:To investigate the impacts of Huayusan decoction on the growth of transplanted liver cancer in nude mice by regulating the microRNA-129-5p/forkhead box C1(FOXC1)axis.Methods:A nude mouse model of transplanted liver cancer was established and grouped into NC group,Huayusan decoction group,Huayusan decoction+inhibitor NC group,and Huayusan decoction+miR-129-5p inhibitor group,with 8 mice in each group.The expression levels of miR-129-5p and FOXC1 mRNA were detected by qRT-PCR,the dual luciferase reporter gene experiment was applied to verify the targeted regulatory relationship between miR-129-5p and FOXC1,the tumor weight and transplanted tumor volume of nude mice were observed and recorded,HE staining was applied to observe the morphology of the transplanted tumor tissue,and TUNEL was applied to detect apoptosis in the tumor tissue,immunohistochemical staining was applied to detect the expression of Ki-67,Cleaved Caspase-3,and FOXC1 in tumor tissue,Western blot was applied to detect the expression of Ki-67,Cleaved Caspase-3,and FOXC1 proteins.[GK4!3]Results:The expression of miR-129-5p in liver cancer cells decreased significantly,and the expression of FOXC1 mRNA increased significantly,especially in HepG2 cells(P<0.05).Dual luciferase reporter gene experiment showed that there was a targeted regulatory relationship between miR-129-5p and FOXC1.Compared with the NC group,the tumor cells in the Huayusan decoction group showed necrosis and increased apoptosis,miR-129-5p and Cleaved Caspase-3 were significantly increased,the mass and volume of the transplanted tumor,and the expression of FOXC1 and Ki-67 were significantly decreased(P<0.05).The inhibitory effect of down-regulation of miR-129-5p reverse Huayusan decoction on hepatocellular carcinoma xenografts(P<0.05).Conclusion:Huayusan decoction may down-regulate the expression of FOXC1 by up-regulating miR-129-5p,thereby inhibiting the growth of transplanted liver cancer.

关 键 词：肝癌 化瘀散水煎剂 微小RNA-129-5p 叉头框蛋白C1 

分 类 号：R735.7[医药卫生—肿瘤]