肝癌细胞自分泌外泌体通过TGF-β1调控自身细胞学行为  

Autocrine exosomes in liver cancer cells are regulated by TGF-β1 autologous cell behavior

作　　者：李文华 王芊文 王小芳[1] 赵彬 耿玉庆 曹明珍 吴向未[2] 陈雪玲[1] LI Wenhua;WANG Qianwen;WANG Xiaofang;ZHAO Bin;GENG Yuqing;CAO Mingzhen;WU Xiangwei;CHEN Xueling(NHC Key Laboratory of Prevention and Treatment of Central Asia High Incidence Diseases,School of Medicine,Shihezi University,Shihezi 832061,China)

机构地区：[1]国家卫生健康委中亚高发病防治重点实验室,石河子大学医学院,石河子832061 [2]石河子大学医学院第一附属医院,石河子832061

出　　处：《中国免疫学杂志》2023年第7期1442-1445,共4页Chinese Journal of Immunology

基　　金：中国医学科学院中央级公益性科研院所基本科研业务费专项资金资助项目(2020-PT330-003);国家自然科学基金地区科学基金项目(82060297);兵团财政科技计划项目区域创新引导计划项目(2021BB006);石河子大学自主支持科研项目(ZZZC202026A)。

摘　　要：目的:研究肝癌Huh7细胞外泌体促进自身转移的机制。方法:培养肝癌Huh7细胞,分离提取外泌体并进行鉴定。将分离的外泌体与Huh7细胞共培养,Transwell比较细胞迁移和侵袭能力;检测外泌体中TGF-β1水平;实时荧光定量PCR和Western blot分别检测TGF-β1、SMAD2/3、EMT相关分子表达。结果:提取的外泌体经Western blot检出CD63、TSG101条带;透射电镜检测出双侧膜结构小体;纳米粒径分析显示颗粒大小集中在100 nm左右。外泌体中检出TGF-β1。外泌体与Huh7细胞共培养,免疫荧光显示外泌体成功进入Huh7细胞。与对照组相比,共培养组细胞迁移和侵袭大幅增加,TGF-β/Smad通路相关分子活化,上皮标志物E-钙黏蛋白、p-Smad7表达下降,间充质标志物-波形蛋白表达升高。结论:肝癌Huh7细胞自分泌的外泌体通过运输TGF-β1上调TGF/Smad通路进而影响肝癌转移,探明了肝癌发展机制,为肝细胞癌治疗提供了新思路。Objective:To study mechanism of liver cancer Huh7 cell exosomes to promote self-metastasis.Methods:Liver can-cer Huh7 cells were cultured,exosomes were isolated,extracted and identified.Isolated exosomes were co-cultured with Huh7 cells,and Transwell was used to compare cell migration and invasion capabilities;TGF-β1 level in exosomes was detected;real-time fluores-cent quantitative PCR and Western blot were used to detect expressions of TGF-β1,SMAD2/3,EMT related molecules.Results:Extracted exosomes was detected CD63 and TSG101 bands by Western blot;transmission electron microscopy detected double-sided membrane structure small bodies;nanometer particle size analysis showed that particle size was concentrated around 100 nm.TGF-β1 was detected in exosomes.Exosomes were co-cultured with Huh7 cells,and immunofluorescence showed that exosomes successfully entered Huh7 cells.Compared with control group,co-culture group has significantly increasing cell migration and invasion,activatal TGF-β/Smad pathway related molecules,decreasing expressions of epithelial marker E-cadherin and p-Smad7,while increasing expression of mesenchymal marker-vimentin.Conclusion:Autocrine exosomes of liver cancer Huh7 cells up-regulate TGF/Smad pathway by transporting TGF-β1 and then affect liver cancer metastasis,further explores mechanism of liver cancer development and provides new ideas for treatment of hepatocellular carcinoma.

关 键 词：肝癌Huh7细胞 外泌体 TGF-Β/SMAD 

分 类 号：R392.12[医药卫生—免疫学]