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Facile preparation of salivary extracellular vesicles for cancer proteomics.

Sun Y, Xia Z, Shang Z, Sun K, Niu X, Qian L, Fan LY, Cao CX, Xiao H - Sci Rep (2016)

Bottom Line: Affinity chromatography column combined with filter system (ACCF) was developed to efficiently remove the high abundant proteins and viscous interferences of saliva.Protein profiling in the SEVs obtained by this strategy was compared with conventional centrifugation method, which demonstrated that about 70% more SEVs proteins could be revealed.Our results demonstrated that SEVs prepared through the developed strategy are valuable samples for proteomics and could serve as a promising liquid biopsy for cancer.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Microbial Metabolism, Laboratory of Analytical Biochemistry and Bioseparation, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China.

ABSTRACT
Extracellular vesicles (EVs) are membrane surrounded structures released by cells, which have been increasingly recognized as mediators of intercellular communication. Recent reports indicate that EVs participate in important biological processes and could serve as potential source for cancer biomarkers. As an attractive EVs source with merit of non-invasiveness, human saliva is a unique medium for clinical diagnostics. Thus, we proposed a facile approach to prepare salivary extracellular vesicles (SEVs). Affinity chromatography column combined with filter system (ACCF) was developed to efficiently remove the high abundant proteins and viscous interferences of saliva. Protein profiling in the SEVs obtained by this strategy was compared with conventional centrifugation method, which demonstrated that about 70% more SEVs proteins could be revealed. To explore its utility for cancer proteomics, we analyzed the proteome of SEVs in lung cancer patients and normal controls. Shotgun proteomic analysis illustrated that 113 and 95 proteins have been identified in cancer group and control group, respectively. Among those 63 proteins that have been consistently discovered only in cancer group, 12 proteins are lung cancer related. Our results demonstrated that SEVs prepared through the developed strategy are valuable samples for proteomics and could serve as a promising liquid biopsy for cancer.

No MeSH data available.


Related in: MedlinePlus

IPA analysis of the candidate SEVs’ protein biomarkers for lung cancer.Number of lung cancer specific proteins involve in cancer related network (A) and cellular movement related network (B). Gray symbols are SEVs proteins identified in this study.
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f7: IPA analysis of the candidate SEVs’ protein biomarkers for lung cancer.Number of lung cancer specific proteins involve in cancer related network (A) and cellular movement related network (B). Gray symbols are SEVs proteins identified in this study.

Mentions: Ingenuity Pathway Analysis (IPA, Redwood City, CA, USA) analysis of these candidate SEVs’ protein biomarkers for lung cancer revealed that 25 of them (about 40%) involved in cancer network (Fig. 7A) and 13 of them (about 20%) were related with cellular movement (Fig. 7B, Table S4). Further, the literature survey showed that out of these 40% proteins, 12 proteins are lung cancer related biomarkers, which includes Annexin family members (Annexin A1, A2, A3, A5, A6, A11), Nitrogen permease regulator 2-like protein (NPRL2), Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), Mucin 1 (MUC1), Prominin-1 (PROM1), Histone H4 (HIST1H4A), and Tumor necrosis factor alpha-induced protein 3 (TNFAIP3).


Facile preparation of salivary extracellular vesicles for cancer proteomics.

Sun Y, Xia Z, Shang Z, Sun K, Niu X, Qian L, Fan LY, Cao CX, Xiao H - Sci Rep (2016)

IPA analysis of the candidate SEVs’ protein biomarkers for lung cancer.Number of lung cancer specific proteins involve in cancer related network (A) and cellular movement related network (B). Gray symbols are SEVs proteins identified in this study.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835767&req=5

f7: IPA analysis of the candidate SEVs’ protein biomarkers for lung cancer.Number of lung cancer specific proteins involve in cancer related network (A) and cellular movement related network (B). Gray symbols are SEVs proteins identified in this study.
Mentions: Ingenuity Pathway Analysis (IPA, Redwood City, CA, USA) analysis of these candidate SEVs’ protein biomarkers for lung cancer revealed that 25 of them (about 40%) involved in cancer network (Fig. 7A) and 13 of them (about 20%) were related with cellular movement (Fig. 7B, Table S4). Further, the literature survey showed that out of these 40% proteins, 12 proteins are lung cancer related biomarkers, which includes Annexin family members (Annexin A1, A2, A3, A5, A6, A11), Nitrogen permease regulator 2-like protein (NPRL2), Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), Mucin 1 (MUC1), Prominin-1 (PROM1), Histone H4 (HIST1H4A), and Tumor necrosis factor alpha-induced protein 3 (TNFAIP3).

Bottom Line: Affinity chromatography column combined with filter system (ACCF) was developed to efficiently remove the high abundant proteins and viscous interferences of saliva.Protein profiling in the SEVs obtained by this strategy was compared with conventional centrifugation method, which demonstrated that about 70% more SEVs proteins could be revealed.Our results demonstrated that SEVs prepared through the developed strategy are valuable samples for proteomics and could serve as a promising liquid biopsy for cancer.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Microbial Metabolism, Laboratory of Analytical Biochemistry and Bioseparation, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China.

ABSTRACT
Extracellular vesicles (EVs) are membrane surrounded structures released by cells, which have been increasingly recognized as mediators of intercellular communication. Recent reports indicate that EVs participate in important biological processes and could serve as potential source for cancer biomarkers. As an attractive EVs source with merit of non-invasiveness, human saliva is a unique medium for clinical diagnostics. Thus, we proposed a facile approach to prepare salivary extracellular vesicles (SEVs). Affinity chromatography column combined with filter system (ACCF) was developed to efficiently remove the high abundant proteins and viscous interferences of saliva. Protein profiling in the SEVs obtained by this strategy was compared with conventional centrifugation method, which demonstrated that about 70% more SEVs proteins could be revealed. To explore its utility for cancer proteomics, we analyzed the proteome of SEVs in lung cancer patients and normal controls. Shotgun proteomic analysis illustrated that 113 and 95 proteins have been identified in cancer group and control group, respectively. Among those 63 proteins that have been consistently discovered only in cancer group, 12 proteins are lung cancer related. Our results demonstrated that SEVs prepared through the developed strategy are valuable samples for proteomics and could serve as a promising liquid biopsy for cancer.

No MeSH data available.


Related in: MedlinePlus