AEG-1对裸鼠肝癌模型中肝癌细胞生长及肺转移的影响  

Effects of AEG-1 on Hepatoma Cell Growth and Lung Metastasis in Nude Mice Hepatoma Model

作　　者：刘谨 周珍珍 Liu Jin;Zhou Zhenzhen(Department of Gastroenterology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China;Department of Gastroenterology,Henan Provincial People’s Hospital,Zhengzhou 450003,China)

机构地区：[1]华中科技大学同济医学院附属同济医院消化内科,武汉430030 [2]河南省人民医院消化内科,郑州450003

出　　处：《华中科技大学学报（医学版）》2023年第2期158-165,共8页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省自然科学基金资助项目(No.2022CFB277)。

摘　　要：目的研究过表达和沉默AEG-1基因对肝癌细胞生长的影响,以及AEG-1基因在调节肝癌细胞定向肺转移中的作用。方法分别以携带过表达AEG-1序列及对照基因序列的慢病毒(lentivirus)转染SMMC-7721肝癌细胞株(SMMC-7721-AEG-1-L;SMMC-7721-control-L);以携带shRNA AEG-1及对照shRNA质粒(plasmid)转染SMMC-7721细胞株(SMMC-7721-shAEG-1-P;SMMC-7721-control-P);使用实时定量PCR和Westen blot检测AEG-1的表达;随后使用荧光素酶基因慢病毒包装颗粒转染上述4种稳定转染细胞株。使用上述细胞株分别建立3种裸鼠肝癌模型:皮下移植瘤模型,原位移植瘤模型和血行播散模型;每种细胞株每一模型5只Balb-c裸鼠,共60只。建立皮下移植瘤模型,观测肿瘤的生长情况并绘制肝细胞肿瘤生长曲线;建立肝癌原位移植瘤和血行播散模型,采用生物发光活体成像技术及组织病理学方法监测造模成功率及肿瘤在肝内、肝外转移情况。结果通过肝癌皮下移植瘤模型可观察到AEG-1过表达组肿瘤体积显著高于对照组,且裸鼠肝脏组织出现弥漫性侵袭转移灶;在肝癌原位移植瘤和血行播散模型中可观察到AEG-1过表达/沉默可以导致肝癌细胞肝内转移、肺转移率和转移灶数量显著增高/降低。结论过表达AEG-1基因可促进肝癌细胞生长以及定向肺转移,沉默AEG-1基因抑制肝癌细胞生长及定向肺转移。Objective To explore effect of overexpression and silencing of AEG 1 gene on the growth of hepatic carcinoma cells,and to confirm whether AEG 1 gene could regulate the targeted lung metastasis of hepatoma cells in vivo.Methods SMMC-7721 hepatoma cell lines(SMMC-7721-AEG-1-L,SMMC-7721-control-L)were transfected with lentivirus carrying AEG-1 sequence and control gene sequence,respectively.SMMC-7721 cell lines(SMMC-7721-shAEG-1-P,SMMC 7721-control-P)were transfected with shRNA AEG 1 and control shRNA plasmid.AEG 1 mRNA expression was detected by real-time quantitative PCR.The four stable transfected cell lines were then transfected with luciferase gene lentivirus package particles.Three types of nude mice hepatoma models were established using the above cell lines:subcutaneous xenograft tumor model,orthotopic xenograft tumor model and hematogenous spread model.Sixty Balb-c nude mice were included,with five mice per cell lines in each group.Tumor growth was observed and the growth curve was plottered in subcutaneous xenograft tumor model.Success rate of modeling as well as intrahepatic and extrahepatic metastasis of tumors were measured by bioluminescent in vivo imaging techniques.Results The tumor volume of the AEG-1 overexpression group was significantly higher than that of the control group,and the liver tissue of nude mice showed diffuse invasion and metastasis in subcutaneous xenograft tumor model.AEG-1 overexpression/silencing was observed in orthotopic xenograft tumor model and hematogenous spread model,which resulted in intrahepatic metastasis,lung metastasis,and significant increase/decrease in number of metastasis foci.Conclusion AEG1 overexpression promotes liver cancer cell growth and lung metastasis.Silencing of AEG 1 gene inhibits the growth of hepatocellular carcinoma cells and lung metastasis.

关 键 词：AEG-1 裸鼠肝癌模型 肝癌肺转移 动物活体成像 

分 类 号：R735.7[医药卫生—肿瘤]