Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Oct 2;90(19):11290-11296.
doi: 10.1021/acs.analchem.8b01831. Epub 2018 Sep 10.

Label-Free Prostate Cancer Detection by Characterization of Extracellular Vesicles Using Raman Spectroscopy

Wooje Lee  1 , Afroditi Nanou  2 , Linda Rikkert  2   3   4 , Frank A W Coumans  4   5 , Cees Otto  2 , Leon W M M Terstappen  2 , Herman L Offerhaus  1
Affiliations

Label-Free Prostate Cancer Detection by Characterization of Extracellular Vesicles Using Raman Spectroscopy

Wooje Lee et al. Anal Chem. .

Abstract

Mammalian cells release extracellular vesicles (EVs) into their microenvironment that travel the entire body along the stream of bodily fluids. EVs contain a wide range of biomolecules. The transported cargo varies depending on the EV origin. Knowledge of the origin and chemical composition of EVs can potentially be used as a biomarker to detect, stage, and monitor diseases. In this paper, we demonstrate the potential of EVs as a prostate cancer biomarker. A Raman optical tweezer was employed to obtain Raman signatures from four types of EV samples, which were red blood cell- and platelet-derived EVs of healthy donors and the prostate cancer cell lines- (PC3 and LNCaP) derived EVs. EVs' Raman spectra could be clearly separated/classified into distinct groups using principal component analysis (PCA) which permits the discrimination of the investigated EV subtypes. These findings may provide new methodology to detect and monitor early stage cancer.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Concentration and size distribution of EV samples measured using NTA. Panels A, B, C, and D represent the NTA result of red blood cell-derived EVs, platelet derived-EVs, PC3-derived EVs, and LNCaP derived-EVs, respectively. The mean size of red blood cell derived EVs is 148 ± 3.7 nm, and its concentration is 0.85 × 108 ± 0.03 × 108 particles/mL. Platelet-derived EVs is 89 ± 4.6 nm and 0.42 × 108 ± 0.02 × 108 particles/mL. PC3-derived EVs is 172 ± 3.7 nm and 1.00 × 108 ± 0.03 × 108 particles/mL. LNCaP-derived EVs is 167 ± 4.4 nm and 1.06 × 108 ± 0.05 × 108 particles/mL.
Figure 2
Figure 2
Transmission electron microscope images of EV subtypes. Arrows point EVs in the figure. (A) red blood cell-derived EVs, (B) platelet-derived EVs, (C) PC3-derived EVs, and (D) LNCaP-derived EVs. Scale bar in each panel is 500 nm.
Figure 3
Figure 3
Raman spectra of each vesicle EV subtypes. Left column of the figure shows the untreated Raman data and curves in the right column shows preprocessed data. For the data processing, background subtraction and baseline correction were conducted. (A and E) Spectrum of red blood cell-derived EVs, (B and F) spectrum of platelet-derived EVs, (C and G) spectrum of PC3-derived EVs, and (D and H) spectrum of LNCaP-derived EVs.
Figure 4
Figure 4
Raman spectra of EV subtypes. The curves are normalized using feature scaling method to enable comparison of the spectra in same scale. Each EV fingerprint shows spectral differences across the fingerprint area. Shaded area shows the main contribution to the separation by PCA. The spectra are vertically segregated for clarity purpose. High-frequency region (right) also shows small discrepancies between EV subtypes.
Figure 5
Figure 5
PCA score plots for the Raman spectra obtained from four EV subtypes (red blood cell-EVs, blue ●; platelet-EVs, green ●; PC3-EVs, red ▲ ; and LNCaP-EVs, pink ▲). Circles represent blood cell-derived EVs and triangles show cancer-derived EVs. Panels A–C were performed on the fingerprint region (400–1800 cm–1), panels D–F were performed on the high-frequency region (2700–3050 cm–1), and panels G–I were performed on the full spectrum (400–3050 cm–1). Panels B and C show good separation among EVs with various cellular origins. Principal component 1 (PComp1), PComp2, and PComp3 account for 87.47%, 5.27%, and 1.36% of total variance, respectively. (A) Score plot for PComp1 and Pcomp2, (B) score plot for PCom1 and PComp3, and (C) score plot for PComp2 and PComp3. In panels B and C, 94.67% and 98% of the data is classified into two categories, respectively, one containing the healthy cell-derived EVs and the other one the prostate cancer-derived EVs.
Figure 6
Figure 6
PCA loading plots corresponding to (A) PComp1, (B) PComp2, and (C) PComp3.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Park J. O.; Choi D. Y.; Choi D. S.; Kim H. J.; Kang J. W.; Jung J. H.; Lee J. H.; Kim J.; Freeman M. R.; Lee K. Y. Proteomics 2013, 13, 2125–2134. 10.1002/pmic.201200323. - DOI - PubMed
    1. van der Pol E.; Böing A. N.; Harrison P.; Sturk A.; Nieuwland R. Pharmacol. Rev. 2012, 64, 676–705. 10.1124/pr.112.005983. - DOI - PubMed
    1. Verma M.; Lam T. K.; Hebert E.; Divi R. L. BMC Clin. Pathol. 2015, 15, 6.10.1186/s12907-015-0005-5. - DOI - PMC - PubMed
    1. Caby M.-P.; Lankar D.; Vincendeau-Scherrer C.; Raposo G.; Bonnerot C. Int. Immunol. 2005, 17, 879–887. 10.1093/intimm/dxh267. - DOI - PubMed
    1. Pisitkun T.; Shen R.-F.; Knepper M. A. Proc. Natl. Acad. Sci. U. S. A. 2004, 101, 13368–13373. 10.1073/pnas.0403453101. - DOI - PMC - PubMed

Publication types