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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

Schematic diagram for EVs isolation from human saliva.
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f1: Schematic diagram for EVs isolation from human saliva.

Mentions: The ACCF system consisted of two parts, the affinity chromatography column (ACC) and membrane filter (F), as shown in Fig. 1. The ACC part was a column prepared in a syringe by packing 0.5 g starch (from potato, Sigma, Shanghai, China) with 3 mL phosphate buffered saline (PBS) (Shanghai Bioscience Co. Ltd), which was adequate for 300 μL saliva sample preparation. The F part was a filter with 5 μm PVDF membrane (Millipore, Billerica, MA, USA).


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)

Schematic diagram for EVs isolation from human saliva.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: Schematic diagram for EVs isolation from human saliva.
Mentions: The ACCF system consisted of two parts, the affinity chromatography column (ACC) and membrane filter (F), as shown in Fig. 1. The ACC part was a column prepared in a syringe by packing 0.5 g starch (from potato, Sigma, Shanghai, China) with 3 mL phosphate buffered saline (PBS) (Shanghai Bioscience Co. Ltd), which was adequate for 300 μL saliva sample preparation. The F part was a filter with 5 μm PVDF membrane (Millipore, Billerica, MA, USA).

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