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CD147、CA9和CD70作为透明细胞肾细胞癌胞外囊泡上肿瘤特异性标志物的特征分析

Characterization of CD147, CA9, and CD70 as Tumor-Specific Markers on Extracellular Vesicles in Clear Cell Renal Cell Carcinoma.

作者信息

Himbert Dirk, Zeuschner Philip, Ayoubian Hiresh, Heinzelmann Joana, Stöckle Michael, Junker Kerstin

机构信息

Department of Urology and Pediatric Urology, Saarland University, 66421 Homburg/Saar, Germany.

Department of Ophthalmology, University Hospital Halle (Saale), Martin-Luther-University Halle-Wittenberg, 06108 Halle/Saale, Germany.

出版信息

Diagnostics (Basel). 2020 Dec 2;10(12):1034. doi: 10.3390/diagnostics10121034.

DOI:10.3390/diagnostics10121034
PMID:33276608
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7761541/
Abstract

Extracellular vesicles (EVs) are secreted by healthy and tumor cells and are involved in cell-cell communication. Tumor-released EVs could represent a new class of biomarkers from liquid biopsies. The aim of this study was to identify tumor-specific EV markers in clear cell renal carcinoma (ccRCC) using cell lines and patient-derived tissue samples. EVs from ccRCC cell lines (786-O, RCC53, Caki1, and Caki2) and patient tissues were isolated via ultracentrifugation. EVs were characterized using transmission electron microscopy, nanoparticle tracking analysis, and Western blotting using exosome and putative tumor markers (epithelial cell adhesion molecule (EpCAM), carbonic anhydrase 9 (CA9), CD70, CD147). The tumor markers were verified using immunohistochemistry. CA9 was expressed in Caki2 cells and EVs, and CD147 was found in the cells and EVs of all tested ccRCC cell lines. In tumor tissues, we found an increased expression of CA9, CD70, and CD147 were increased in cell lysates and EV fractions compared to normal tissues. In contrast, EpCAM was heterogeneously expressed in tumor samples and positive in normal tissue. To conclude, we developed an effective technique to isolate EVs directly from human tissue samples with high purity and high concentration. In contrast to EpCAM, CA9, CD70, and CD147 could represent promising markers to identify tumor-specific EVs in ccRCC.

摘要

细胞外囊泡（EVs）由健康细胞和肿瘤细胞分泌，参与细胞间通讯。肿瘤释放的细胞外囊泡可能代表液体活检中的一类新型生物标志物。本研究的目的是利用细胞系和患者来源的组织样本，鉴定透明细胞肾细胞癌（ccRCC）中肿瘤特异性的细胞外囊泡标志物。通过超速离心从ccRCC细胞系（786 - O、RCC53、Caki1和Caki2）和患者组织中分离细胞外囊泡。使用透射电子显微镜、纳米颗粒跟踪分析以及利用外泌体和假定的肿瘤标志物（上皮细胞粘附分子（EpCAM）、碳酸酐酶9（CA9）、CD70、CD147）进行蛋白质免疫印迹对细胞外囊泡进行表征。使用免疫组织化学验证肿瘤标志物。CA9在Caki2细胞和细胞外囊泡中表达，并且在所有测试的ccRCC细胞系的细胞和细胞外囊泡中均发现有CD147。在肿瘤组织中，我们发现与正常组织相比，细胞裂解物和细胞外囊泡组分中CA9、CD70和CD147的表达增加。相比之下，EpCAM在肿瘤样本中表达不均一，在正常组织中呈阳性。总之，我们开发了一种有效的技术，可直接从人体组织样本中分离出高纯度和高浓度的细胞外囊泡。与EpCAM不同，CA9、CD70和CD147可能是鉴定ccRCC中肿瘤特异性细胞外囊泡的有前景的标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/7761541/14c8c7c903e6/diagnostics-10-01034-g010.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/7761541/11ca33696c9e/diagnostics-10-01034-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/7761541/b677707e1fb1/diagnostics-10-01034-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/7761541/a35de9047086/diagnostics-10-01034-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/7761541/14c8c7c903e6/diagnostics-10-01034-g010.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/7761541/3a2858312cd0/diagnostics-10-01034-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/7761541/4b121587d0fc/diagnostics-10-01034-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/7761541/11ca33696c9e/diagnostics-10-01034-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/7761541/b677707e1fb1/diagnostics-10-01034-g008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ec8/7761541/14c8c7c903e6/diagnostics-10-01034-g010.jpg

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