【作者】 王晨昱；

【导师】 李敏；

【作者基本信息】 兰州大学 ， 基础医学， 2025， 博士

【摘要】 背景肿瘤血道转移是患者预后不良的重要原因。血道转移是一个多步骤级联的过程,癌细胞入侵发育不良和高渗漏性的血管进入循环,诱导内皮屏障功能变化,完成跨内皮外渗并定植是导致肿瘤血道转移的关键,但调控转移的机制尚不清楚。因靶器官组织血管内皮细胞发育完全,其渗透性有限,循环血液中的癌细胞是如何黏附远端靶器官的内皮细胞并跨内皮迁移?哪些黏附配体介导了癌细胞与内皮细胞的黏附?这些关键的问题尚未解决。血管性血友病因子(von Willebrand factor,VWF)是血液中的黏附蛋白,我们前期研究发现胃癌患者血液中VWF水平显著升高,且与低分化和转移正相关;肿瘤源性VWF可介导癌细胞间黏附及癌细胞与血小板的相互作用。血液中的VWF作为重要的黏附配体,也是内皮活化或损伤的标志,此时内皮细胞分泌超大型多聚体VWF在内皮表面组装成超黏附VWF串(VWF strings),或致血浆VWF多聚体增加。那么VWF是如何介导循环血液中的癌细胞与内皮细胞黏附而外渗的,还需进一步研究。细胞外囊泡(Extracellular vesicles,EVs)作为细胞间信息交流的载体,有研究报道肿瘤源性EVs可促进恶性肿瘤的增殖、侵袭和转移。我们前期研究发现,胃癌患者血浆升高的胃癌源性细胞外囊泡(Gastric cancer cell-derived EVs,gc EVs)可通过增加血管内皮通透性,增强肿瘤细胞跨内皮迁移并促进胃癌血道转移,但是gc EVs是如何黏附内皮细胞而介导内皮损伤的机制还不清楚。在创伤性脑损伤的小鼠血浆中,存在高黏附VWF绑定的EVs促进凝血并显著增加内皮通透性。我们前期也发现在胃癌患者血浆中存在VWF绑定EVs的形式,且VWF黏附gc EVs水平升高,这些高黏附VWF是否能协同EVs促进胃癌转移。因此我们推测:1)高黏附VWF作为重要的黏附配体,在循环血液中绑定gc EVs介导其与内皮细胞相互作用,影响内皮屏障功能并促进胃癌转移;2)VWF-gc EV的绑定与整合素分子相关;3)临床胃癌患者血浆存在高黏附VWF绑定的EVs与肿瘤进展相关;4)降低VWF的黏附活性和或清除EVs能够减少胃癌的血道转移。目的本研究旨在探讨胃癌中高黏附VWF作为关键的耦联因子,将循环中的gc EVs捕获至内皮细胞,GcEVs以高黏附VWF介导方式增强与内皮细胞黏附,增加内皮屏障通透性,促进胃癌细胞黏附并迁移、促进血道转移;揭示ADAMTS-13及乳凝集素对减少胃癌血道转移的作用;探讨整合素αVβ3介导高黏附VWF绑定gc EVs的分子机制;探讨胃癌患者循环中高黏附VWF绑定EVs的水平及与临床病理因素的相关性和诊断价值。方法1.检测高黏附VWF绑定gc EVs对胃癌血道转移的影响。建立615小鼠MFC细胞皮下移植瘤模型,分离、提取、鉴定小鼠胃癌gc EVs。小鼠胃癌肺转移模型,分别或联合尾静脉注射gc EVs、rh VWF、VWF结合的gc EVs、乳凝集素和ADAMTS-13,应用ELISA、流式细胞术检测模型鼠血浆中VWF:Ag、VWF:CB、ADAMTS-13:Ag、VWF~+/PS~+EVs及Ep CAM~+/PS~+EVs的水平与肺转移的关系。阐明高黏附VWF绑定gc EVs体内对胃癌肺转移的影响,降低VWF黏附活性及清除EVs能抑制胃癌血道转移。2.检测胃癌细胞外囊泡诱导内皮激活与黏附功能的改变。分离、提取、鉴定人胃癌细胞系gc EVs,收集经gc EVs处理的内皮细胞培养上清,ELISA检测VWF:Ag、VWF:CB,流式细胞术检测VWF~+EVs、e EVs,蛋白芯片检测细胞因子水平;应用免疫荧光、WB检测内皮高黏附VWF串及与gc EVs黏附、内吞情况,以及内皮骨架蛋白、相关黏附蛋白的表达变化。3.检测高黏附VWF绑定gc EVs黏附内皮、增加内皮屏障通透性。应用静态及动态黏附实验,测试rh VWF在介导gc EVs与内皮黏附中的作用。应用Transwell内皮屏障模型,测量跨内皮电阻及异硫氰酸荧光素-右旋糖酐的渗漏,验证rh VWF协同gc EVs促进内皮屏障渗漏的作用;应用VWF阻断抗体反向验证抑制VWF黏附活性对其介导的gc EVs诱导内皮屏障功能的影响。4.检测高黏附VWF绑定gc EVs对胃癌细胞侵袭迁移的影响。收集gc EVs处理内皮细胞后的条件培养基,检测VWF绑定的gc EVs介导胃癌细胞迁移及侵袭的作用。应用免疫荧光及Transwell模型验证rh VWF协同gc EVs促进HGC-27和AGS胃癌细胞黏附内皮及跨内皮迁移。5.高黏附VWF绑定gc EVs的分子机制研究。应用TCGA数据库分析胃癌整合素αV与β3的表达及其生物学功能;WB及流式细胞术检测胃癌细胞及gc EVs的整合素αVβ3表达水平;体外应用整合素αVβ3拮抗剂西仑吉肽验证对高黏附VWF绑定gc EVs的拮抗作用,及其影响胃癌细胞黏附内皮的作用。6.检测胃癌组织中高表达VWF与内皮细胞相关性,研究临床患者血浆中gc EVs与激活内皮释放高黏附VWF绑定EVs的相关性。分析TCGA数据库胃癌的VWF表达及与临床病理因素的关系;应用肿瘤免疫评估资源的“Immune-Gene”,分析VWF表达与胃癌相关内皮细胞的相关性;利用单细胞数据库TISCH分析VWF在不同细胞的表达水平;免疫组化检测胃癌组织VWF的表达情况。流式细胞术检测胃癌患者血浆中gc EVs、VWF~+EVs、VWF~+gc EVs及e EVs水平及相关性分析。7.研究胃癌组织中整合素αVβ3表达与肿瘤进展的关系,临床患者血浆中高黏附VWF绑定EVs的水平与胃癌进展及诊断价值。分析TCGA胃癌的ITGAV表达与临床病理因素的关系;免疫组化检测胃癌组织ITGAV的表达与临床病理因素的关系。流式检测患者血浆中αVβ3~+gc EVs水平与VWF~+EVs的相关性;分析各类EVs与临床病理因素的关系,阐明各类EVs在胃癌进展中作用;验证联合检测EVs在胃癌中的诊断价值。结果1.胃癌皮下荷瘤及尾静脉肺转移模型鼠血浆中gc EVs水平均升高,血浆VWF:Ag水平及黏附活性均显著增加。肺转移模型注射gc EVs的小鼠肺转移率增加;同时血浆gc EVs、VWF:Ag、VWF黏附活性及ADAMTS-13:Ag与VWF:Ag的比例显著降低;VWF~+/PS~+EVs及VWF~+/CD62E~+EVs比例增高。注射高黏附rh VWF与gc EVs的小鼠具有更高的肺转移率;小鼠肺组织中VWF主要表达在间质血管内皮细胞,癌组织弱表达VWF,注射gc EVs的小鼠肺间质血管内皮表达VWF增强。应用ADAMTS-13联合Lactadherin可显著降低血浆VWF:Ag水平、黏附活性及PS~+EVs的水平,减少肺转移。2.胃癌细胞外囊泡可激活内皮释放VWF:Ag及VWF~+e EVs至上清,同时内皮细胞胞质颗粒和膜表面广泛表达VWF串且黏附gc EVs并共定位。GcEVs诱导内皮激活释放各类细胞因子,与促炎和增加血管通透性有关,并引起内皮骨架蛋白变化。高黏附rh VWF促进gc EVs诱导的黏附蛋白GP1BA与VCAM-1的表达上调,VWF阻断抗体可减弱此作用。3.高黏附rh VWF促进静态与流体剪切力作用下gc EVs与内皮细胞黏附,VWF阻断抗体可抑制黏附。高黏附rh VWF协同gc EVs促进内皮通透性增加,VWF阻断抗体可拮抗此作用。4.高黏附rh VWF或经gc EVs刺激的内皮条件培养基可协同gc EVs促进胃癌细胞的迁移和侵袭;高黏附rh VWF协同gc EVs促进胃癌细胞黏附内皮细胞并跨内皮迁移,VWF阻断抗体可拮抗此作用。5.TCGA数据库胃癌中整合素ITGAV与ITGB3表达显著上调;高表达ITGAV患者的总生存率与无病生存期显著降低。筛选差异表达基因,高表达ITGAV与ITGB3主要的基因功能与细胞外基质结构组成及ECM受体互作有关。胃癌细胞外囊泡膜富集整合素αVβ3,西仑吉肽可抑制整合素αVβ3表达影响VWF绑定gc EVs及黏附内皮细胞;西仑吉肽可抑制胃癌细胞黏附rh VWF,可拮抗gc EVs促进的胃癌细胞黏附内皮的作用。6.TCGA数据库胃癌中VWF表达水平显著升高,与胃癌较晚的分期相关;高表达VWF的胃癌患者总生存率降低;胃癌中VWF的表达与癌相关内皮细胞数量呈正相关。癌组织血管内皮见大量VWF阳性表达;癌细胞可部分表达VWF,分化越低的癌细胞VWF表达较强。胃癌患者血浆中gc EVs、VWF~+/PS~+EVs、e EVs、VWF~+gc EVs水平显著升高;VWF~+/PS~+EVs水平与gc EVs、e EVs水平显著正相关。7.TCGA数据库胃癌中ITGAV表达与T分期、N分期、p TNM及病理分级有关;分化低的胃癌组织ITGAV表达较强。胃癌患者血浆中αVβ3~+/PS~+EVs水平显著升高,与VWF~+/PS~+EVs正相关;VWF~+gc EVs水平αVβ3~+gc EVs正相关;血浆各类EVs水平随着癌组织浸润、淋巴结转移、病理分级各组间均有升高趋势。联合检测PS阳性的gc EVs、VWF~+EVs、VWF~+gc EVs及αVβ3~+EVs水平,具有更高的AUC值。结论1.胃癌荷瘤小鼠体内存在的高黏附VWF与gc EVs诱导内皮活化有关,胃癌源性细胞外囊泡引起小鼠获得性ADAMTS-13缺乏,使高黏附VWF在血浆及循环EVs积聚,促进了胃癌肺转移;ADAMTS-13剪切高黏附VWF并联合乳凝集素清除循环EVs对抑制胃癌转移起叠加作用。2.胃癌细胞外囊泡激活内皮释放高黏附VWF,高黏附VWF绑定gc EVs增强内皮屏障黏附功能并导致通透性升高,增强肿瘤细胞的侵袭迁移能力。通过靶向VWF可起一定的抑制作用。3.整合素拮抗剂西仑吉肽抑制高黏附VWF介导的gc EVs黏附内皮,从而减少胃癌细胞黏附经gc EVs诱导活化的内皮。提示整合素αVβ3是高黏附VWF绑定gc EVs的特异性受体。4.胃癌患者体内高黏附VWF与内皮细胞数量及gc EVs诱导的内皮活化呈正相关;肿瘤进展中,整合素αVβ3~+的胃癌细胞外囊泡与血浆中高黏附VWF绑定EVs呈正相关;联合检测gc EVs、VWF~+EVs、VWF~+gc EVs及αVβ3~+EVs在胃癌中具有更高的诊断价值。更多还原

【Abstract】 Background The hematogenous metastasis of cancers,is a multistep process,significantly contributes to poor clinical outcomes in patients.During which cancer cells are released into the circulation through the poorly developed and“leaky”vessels of the cancer tissue niche.When in the circulation,cancer cells adhere to the endothelium of selective organs,changes in endothelial barrier functions locally,and transmigrate through the endothelium to form new cancer loci,which is key to tumor hematogenous metastasis.However,the mechanisms of regulating metastasis are still unclear.How cancer cells in rapidly circulating blood adhere to and transmigrate through the endothelium of a remote organ,where the endothelium is fully developed and has limited permeability,and which adhesive ligands mediate the adhesion of cancer cells to the endothelium.These critical questions remain unresolved.Previous studies have found that the levels of von Willebrand factor(VWF)are significantly increased in blood samples of patients with GCs,are associated with poorly differentiated and metastatic cancers of patients with GCs.Tumor-derived VWF can mediate adhesion between cancer cells and interactions between cancer cells and platelets.Among adhesive ligands,VWF serves as a biomarker of endothelial activation or injury,and endothelial cells secrete ultra-large VWF multimers which assemble into hyperadhesive VWF strings on the endothelial surface,or lead to an increase in plasma VWF multimers.How VWF mediates the adhesion of cancer cells to the endothelium in the circulation and extravasation needs further research.Extracellular vesicles(EVs)serve as carriers of intercellular information exchange,and have been reported to promote the proliferation,invasion and metastasis of malignant tumors.Our previous studies have found that elevated gastric cancer cell-derived EVs(gc EVs)in blood samples of patients with GCs,can elevate endothelial permeability and transendothelial migration of cancer cells to promote GC metastasis.However,the mechanism by which gc EVs adhere to the endothelium and mediate endothelial injury is still unclear.In the plasma of mice with traumatic brain injury,the presence of hyperadhesive VWF-bound EVs promote coagulation and significantly increase endothelial permeability.We also found that the levels of VWF adhering to gc EVs is elevated in blood samples of patients with GCs.Whether these hyperadhesive VWF synergize with EVs to promote the progression and metastasis of gastric cancer.We therefore hypothesize that:(1)Hyperadhesive VWF,as an important adhesive ligand,binds to gc EVs in the circulation to mediate their interaction with endothelial cells,affects endothelial barrier function,through which cancer cells metastasize.(2)The binding of VWF to gc EVs is related to integrin molecules.(3)The presence of hyperadhesive VWF-bound EVs in blood samples of patients with GCs,is associated with tumor progression.(4)Reducing VWF hyperadhesive activity and clearing EVs can prevent the hematogenous metastasis of gastric cancer.Objective This study aims to investigate whether hyperadhesive VWF,serves as the key tethering factor to capture circulating gc EVs to endothelial cells,where gc EVs enhance adhesion to endothelial cells and increase endothelial barrier permeability in a hyperadhesive VWF-mediated manner,thereby promoting transendothelial migration of cancer cells to promote GC metastasis.It also seeks to elucidate the roles of ADAMTS-13 and lactadherin in reducing the hematogenous metastasis of gastric cancer.To investigate the molecular mechanisms of integrinαVβ3 in mediating the adhesion of hyperadhesive VWF to gc EVs.To investigate the levels of hyperadhesive VWF-bound EVs and their correlation with clinical pathological factors and diagnostic value in patients with GCs.Methods1.To detect the impact of hyperadhesive VWF binding to gc EVs on hematogenous metastasis of gastric cancer.A subcutaneous xenograft tumor model of 615 mice was established,and mouse gc EVs were isolated,extracted,and identified.Mouse model of gastric cancer pulmonary metastasis was established.Mice were intravenously injected with gc EVs,rh VWF,VWF-bound gc EVs,galectin,and ADAMTS-13,either separately or in combination.ELISA and flow cytometry were applied to detect the levels of VWF:Ag,VWF:CB,ADAMTS-13:Ag,VWF~+/PS~+EVs and Ep CAM~+/PS~+EVs in the plasma of tumor-bearing mice,and their correlation with pulmonary metastasis.The study elucidated the effect of hyperadhesive VWF-bound gc EVs on gastric cancer pulmonary metastasis in vivo,and demonstrated that reducing VWF hyperadhesive activity and removing EVs could inhibit the hematogenous metastasis of gastric cancer.2.To detect the changes in endothelial activation and adhesion functions induced by gc EVs.GcEVs were isolated,extracted and identified from human gastric cancer cell lines.The culture supernatant of endothelial cells treated with gc EVs was collected,subsequently,ELISA and flow cytometry were used to detect VWF:Ag,VWF:CB,VWF~+EVs and e EVs,and cytokine levels were detected by protein chip.Immunofluorescence and western blotting were used to detect the expression of endothelial hyperadhesive VWF strings and their adhesion and internalization with gc EVs,as well as changes in the expression of endothelial cytoskeletal protein and related adhesion proteins.3.Detection of hyperadhesive VWF binding gc EVs to adhere to endothelium and increase barrier permeability.The role of rh VWF in mediating the adhesion of gc EVs to endothelium were conducted by static and dynamic adhesion experiments.The transendothelial resistance and fluorescein isothiocyanate-dextran leakage were measured by transwell endothelial barrier model to verify the effect of rh VWF combined with gc EVs on endothelial barrier leakage.VWF-blocking antibodies were applied to reverse-verify the effect of reducing VWF-bound gc EVs in mediating EV-induced endothelial barrier function.4.To detect the effect of hyperadhesive VWF-bound gc EVs on the invasion and migrationof gastric cancer cells.Conditioned medium was collected from gc EVs-treated endothelial cells to detect the role of VWF-bound gc EVs in mediating the migration and invasion of gastric cancer cells.Immunofluorescence and the transwell model were applied to verify that rh VWF synergized with gc EVs promoted the adhesion to endothelium and transendothelial migration of HGC-27 and AGS gastric cancer cells.5.The molecular mechanisms of hyperadhesive VWF binding gc EVs.The expression of integrinαV andβ3 in gastric cancer and their biological functions were analyzed using the TCGA database.The expression of integrinαVβ3 in gastric cancer cells and gc EVs was detected by western blotting and flow cytometry.In vitro,the integrinαVβ3 antagonist cilengitide was used to verify the antagonistic effect on hyperadhesive VWF-bound gc EVs and its impact on gastric cancer cell adhesion to endothelium.6.To detect the correlation between overexpressed VWF and endothelium in gastric cancer tissues,and to study the correlation between gc EVs and activated endothelial release of hyperadhesive VWF-bound EVs in blood samples of patients with GCs.The expression of VWF in gastric cancer and its correlation with clinical pathological factors were analyzed using the TCGA database as the main database.The correlation between VWF expression and gastric cancer-related endothelium was analyzed using the“Immune-Gene”from the Tumor Immune Estimation Resource.The expression levels of VWF in different cells were analyzed using the single-cell database TISCH.The expression of VWF in gastric cancer tissues was detected by immunohistochemistry.Flow cytometry was used to detect the levels of gc EVs,VWF~+EVs,VWF~+gc EVs and e EVs in blood samples of patients with GCs and analyzed their correlation.7.To study the relationship between the expression of integrinαVβ3 and tumor progression in gastric cancer tissues,and correlation analysis of hyperadhesive VWF-bound EVs with clinical pathological factors and diagnostic value.The relationship between ITGAV expression in GC and clinical pathological factors was analyzed using the TCGA database.The expression of ITGAV in gastric cancer tissues were detected by immunohistochemistry and its correlation were analyzed with clinical pathological factors.Flow cytometry was used to detect the levels ofαVβ3~+gc EVs in patient plasma and their correlation with VWF~+EVs.To analyze the relationship between various EVs and clinical pathological factors,and to clarify the role of various EVs in the progression of gastric cancer.To verify the diagnostic value of combined detection of EVs in gastric cancer.Results1.The levels of plasma gc EVs,VWF:Ag and adhesive activity were significantly increased in both subcutaneous tumor-bearing and pulmonary metastasis model mice of gastric cancer.Mice injected with MFC cells and gc EVs through the tail had a higher rate of pulmonary metastasis,concurrently,the levels of plasma gc EVs,VWF:Ag,VWF adhesive activity,VWF~+/PS~+EVs and VWF~+/CD62E~+EVs were elevated in these mice,and the ADAMTS-13-to-VWF ratio was significantly reduced in mice receiving MFC alone and with gc EVs.The rate of pulmonary metastasis increased drastically in mice receiving MFC together with both rh VWF and gc EVs.VWF was mainly expressed in the interstitial vascular endothelium of lung tissues in mice,and VWF was weakly expressed in cancer cells.VWF expression enhanced in vascular endothelium when mice injected MFC together gc EVs.The application of ADAMTS-13 combined with Lactadherin can significantly reduce plasma VWF:Ag level,adhesion activity and PS~+EVs level,and reduce lung metastasis.2.Endothelial cells activated by gc EVs released VWF and shed endothelial VWF~+EVs into the supernatant,while the expression of VWF not only in intracellular Weibel-Palade bodies but also on the surface,where string-like VWF fibrils were formed,which adhere to and colocalize with gc EVs.GcEVs induced endothelial activation to release various cytokines,which were associated with proinflammation and increased endothelial permeability,causing changes in endothelial cytoskeletal proteins.Hyperadhesive VWF promoted gc EVs-induced upregulation of the expression of the adhesion proteins GP1BA and VCAM-1,which could be attenuated by VWF-blocking antibody.3.Hyperadhesive VWF promoted the adhesion of gc EVs to endothelial cells under static and fluid shear stress conditions,and hyperadhesive VWF synergized with gc EVs to promote endothelial permeability,which could be attenuated by VWF-blocking antibody.4.Hyperadhesive VWF,or endothelial-conditioned medium stimulated by gc EVs synergized with gc EVs to promote the migration and invasion of gastric cancer cells.Hyperadhesive rh VWF could synergize with gc EVs to promote the adhesion and transendothelial migration of gastric cancer cells to endothelial cells,which could be antagonized by VWF blocking antibody.5.The expression of integrins ITGAV and ITGB3 was significantly up-regulated in gastric cancer in the TCGA database.The overall survival and disease-free survival probability of patients with high levels of ITGAV m RNA was significantly lower than patients with low levels of ITGAV m RNA.Differential expression gene analysis reveals that the main gene functions associated with high ITGAV and ITGB3expression were related to the structural composition of the extracellular matrix and ECM receptor interaction.GcEVs were enriched with integrinαVβ3 on their membranes.By using the integrinαVβ3 inhibitor cilengitide,which could inhibit the the expression of integrinαVβ3 affecting the binding of VWF to gc EVs,and reduce adhesion to endothelial cells.Cilengitide could inhibit gastric cancer cell adhesion to rh VWF and the endothelium activated by gc EVs.6.The TCGA database showed that gastric cancer tissue expressed significantly higher levels of VWF m RNA than noncancerous tissue,which was related to the later stage of gastric cancer.The overall survival probability of patients with high levels of VWF m RNA was significantly lower than patients with low levels of VWF m RNA.VWF is mainly expressed in the interstitial vascular endothelial cells of cancer tissues,and cancer cells can partially express VWF,and more poorly differentiated cancer cells have stronger VWF expression.The levels of gc EVs,VWF~+/PS~+EVs,e EVs and VWF~+gc EVs were significantly elevated in blood samples of patients with GCs,and the level of VWF~+/PS~+EVs was significantly positively correlated with the levels of gc EVs and e EVs.7.The TCGA database showed that ITGAV expression in gastric cancer was associated with T stage,N stage,p TNM and pathological grade.ITGAV expression was stronger in poorly differentiated gastric cancer tissues.The level ofαVβ3~+/PS~+EVs was significantly elevated and positively correlated with VWF~+/PS~+EVs in blood samples of patients with GCs.The level of VWF~+gc EVs was positively correlated withαVβ3~+gc EVs.The higher levels of various EVs in plasma were found in cancer invasion,lymph node metastasis and poorly differentiated gastric cancer.Combined detection of PS-positive gc EVs,VWF~+EVs,VWF~+gc EVs,andαVβ3~+EVs levels had higher AUC values.Conclusion1.The presence of hyperadhesive VWF in gastric cancer-bearing mice is associated with gc EVs-induced endothelial activation.Gastric cancer-derived extracellular vesicles cause acquired ADAMTS-13 deficiency in mice,leading to the accumulation of hyperadhesive VWF in plasma and circulating EVs,which promotes pulmonary metastasis of gastric cancer.The combined effect of ADAMTS-13 cleaving hyperadhesive VWF and lactadherin clearing circulating EVs plays a synergistic role in inhibiting gastric cancer metastasis.2.Gastric cancer extracellular vesicles activate endothelial cells to release hyperadhesive VWF.Hyperadhesive VWF binding to gc EVs enhances endothelial barrier adhesion function and leads to increased permeability,increasing the invasive and migratory capacity of cancer cells.By targeting VWF can play a certain inhibitory effect.3.The integrin antagonist cilengitide inhibits the adhesion of gc EVs to endothelial cells mediated by hyperadhesive VWF,thereby reducing the adhesion of gastric cancer cells to endothelial cells activated by gc EVs.This suggests that integrinαVβ3 is a specific receptor for hyperadhesive VWF binding to gc EVs.4.Hyperadhesive VWF is positively correlated with the number of endothelial cells and endothelial activation induced by gc EVs in gastric cancer patients.IntegrinαVβ3~+gc EVs are positively correlated with with hyperadhesive VWF-bound EVs in plasma during tumor progression.Combined detection of gc EVs,VWF~+EVs,VWF~+gc EVs andαVβ3~+EVs has higher diagnostic value in gastric cancer.更多还原