Abstract:
AIM:To develop a novel method for the rapid and efficient extraction of exosomes secreted by tumor cells. METHODS:Unlike the traditional extraction method, the supernatants of cell cultures were concentrated, and the exosomes were isolated promptly and effectively using a novel nanomaterial called ExoQuick. Coomassie brilliant blue staining was used for protein quantification, and the morphology of the exosomes extracted by both methods was visualized by transmission electron microscopy. Exosome marker proteins were detected by Western blot analysis. Two potential hepatoma-associated proteins, tissue transglutaminase 2 (TGM2) and annexin A2, were analyzed. RESULTS:The exosomes separated by the new extraction assay based on the nanomaterial were disc-shaped, intact vesicles with lipid bilayer membranes. They were approximately 30-100 nm in diameter, which is similar to the diameter of exosomes isolated by the traditional method. The protein concentration of exosomes extracted by the new method was approximately 780 μg/10(8) cells, and therefore, it was 19 times higher than that of exosomes extracted in the traditional manner. There were differences between the total proteins of Huh-7 cells and the exosomal proteins. Typical exosome proteins, such as the transmembrane protein CD63 and heat shock protein 70, were confirmed. Two potential hepatoma-associated proteins were also identified. TGM2 was first found to exist in the exosomes of human liver cancer cells, but annexin A2 was not secreted into exosomes. CONCLUSION:The new extraction method based on the nanomaterial is quick and efficient. The cancer-associated protein TGM2 can be secreted through an exosome-mediated non-classical secretion pathway, and it may be a valuable tumor marker.
journal_name
World J Gastroenteroljournal_title
World journal of gastroenterologyauthors
Zhu L,Qu XH,Sun YL,Qian YM,Zhao XHdoi
10.3748/wjg.v20.i21.6651subject
Has Abstractpub_date
2014-06-07 00:00:00pages
6651-7issue
21eissn
1007-9327issn
2219-2840journal_volume
20pub_type
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更新日期:2003-11-01 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章
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pub_type: 杂志文章
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
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pub_type: 杂志文章
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pub_type: 杂志文章
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pub_type: 临床试验,杂志文章
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v16.i18.2302
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pub_type: 杂志文章,评审
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更新日期:2007-12-21 00:00:00
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