摘要
目的探讨血管包绕的肿瘤细胞巢(VETC)阳性型肝细胞癌癌巢转移的分子机制。方法收集72例肝细胞肝癌标本,通过免疫组化CD34(血管内皮细胞标记蛋白)染色观察VETC癌巢在原发灶、胆管癌栓、门静脉癌栓中的形态学表现,观察VETC癌巢血行转移的特点。利用生物信息学预测与VETC癌巢形成密切相关的分子蛋白,用免疫组化检测血管生成素-2(Ang-2)、整合素α5(integrinα5)、整合素β1(integrinβ1)及环氧合酶-2(COX-2)蛋白在肝癌中的表达,用Transwell细胞迁移实验检测Ang-2/integrinα5β1蛋白对内皮细胞、肝癌细胞迁移能力的影响。Western印迹法检测Ang-2/integrinα5β1蛋白对黏着斑激酶(FAK)蛋白活性的影响。结果在收集的肝癌标本中,VETC(+)肝癌27例(27/72),其中3例存在胆管癌栓,5例存在门静脉癌栓,3例同时存在胆管癌栓、门静脉癌栓。VETC(+)肝癌能够以癌巢的形式发生门静脉、胆管及肝内转移,并保留完整的VETC癌巢结构。Ang-2、integrinα5、integrinβ1可过表达于VETC(+)肝癌癌巢的肿瘤细胞、内皮细胞;COX-2仅高表达于VETC(+)肝癌癌巢的肿瘤细胞。Ang-2可促进肝癌细胞(121±12比186±11,P<0.01)、内皮细胞的迁移(81±7比163±14,P<0.01)。整合素α5β1的激活拮抗剂ATN-161可有效降低Ang-2促肝癌细胞(185±10比135±9,P<0.05)、内皮细胞(156±14比103±6,P<0.05)的迁移能力。ATN-161可显著降低Ang-2对肝癌细胞、内皮细胞FAK蛋白的磷酸化激活。结论VETC(+)肝癌可以癌巢形式发生整体转移,具有特异性的调控蛋白,Ang-2/α5β1/FAK是VETC(+)肝癌癌巢转移治疗中的潜在蛋白靶点。
Objective To investigate the molecular mechanism of nest metastasis in blood vessels encapsulated by tumor clusters(VETC)positive hepatocellular carcinoma(HCC).Methods A total of 72 paraffin embedded HCC tissue samples were collected.Immunohistochemistry staining with CD34(vascular endothelial cell marker protein)was used to observe the morphological manifestations of VETC cancer nests in primary tumors,bile duct cancerous thrombi and portal vein cancerous thrombi,and to study the characteristics of hematogenous metastasis of VETC cancer nests.Bioinformatics was used to predict the key proteins closely related to VETC cancer nest formation.Immunohistochemistry was used to detect the expression of angiogenin-2(Ang-2),integrinα5,Integrinβ1,and cyclooxygenase-2(COX-2)proteins in HCC.Transwell cell migration assay was used to detect the effect of Ang-2/integrinα5β1 protein on the migration ability of endothelial cells and HCC cells.Western blotting was used to detect the effect of Ang-2/integrinα5β1 protein on the activity of focal adhesion kinase(FAK)protein.Results Of the collected HCC specimens,27 cases(27/72)were VETC(+),including 3 cases with biliary duct cancerous thrombus,5 cases with portal vein cancerous thrombus,and 3 cases with both biliary duct cancerous thrombus and portal vein cancerous thrombus.VETC(+)HCC could metastasize to portal vein,bile duct,and liver in the form of cancer nest,and the nests retain their intact structure.Ang-2,integrinα5 and integrinβ1 were overexpressed in tumor cells and endothelial cells of VETC(+)HCC nests,while COX-2 was only overexpressed in tumor cells of VETC(+)HCC nest.Ang-2 could promote the migration of HCC cell[(121±12)vs(186±11),P<0.01]and endothelial cells[(81±7)vs(163±14),P<0.01].Integrinα5β1 activation antagonist ATN-161 could significantly block the ability of Ang-2 to promote the migration of HCC cells[(185±10)vs(135±9),P<0.05]and endothelial cells[(156±14)vs(103±6),P<0.05].ATN-161 could significantly block the phosphorylation of FAK in HCC and endothelial cells induced by Ang-2.Conclusions VETC(+)HCC could metastasize as a whole in a nested form,and possesses a specific regulatory protein.Ang-2/α5β1/FAK might be potential protein targets in the treatment of VETC(+)HCC nest-type metastasis.
作者
董小锋
钟敬涛
刘天奇
陈元元
唐耘天
杨建荣
Dong Xiaofeng;Zhong Jingtao;Liu Tianqi;Chen Yuanyuan;Tang Yuntian;Yang Jianrong(Department of Hepatobiliary,Pancreas and Spleen Surgery,the People′s Hospital of Guangxi Zhuang Autonomous Region,Nanning 530021,China;Department of Hepatobiliary Surgery,Shandong Cancer Hospital and Institute,Shandong First Medical University and Shandong Academy of Medical Sciences,Jinan 250117,China)
出处
《中华医学杂志》
CAS
CSCD
北大核心
2021年第9期654-660,共7页
National Medical Journal of China
基金
国家自然科学基金 (81560406)
广西自然科学基金 (2018GXNSFAA050118)
广西区域性高发肿瘤早期防治研究重点实验室开放课题 (GXK201604)。
关键词
癌
肝细胞
血管包绕的肿瘤细胞巢
血管生成素-2
整合素
Carcinoma,hepatocellular
Vessels encapsulated by tumor clusters
Angiopoietin-2
Integrin
