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ICTMI 2017 pp 163-174 | Cite as

Probable Role of Non-exosomal Extracellular Vesicles in Colorectal Cancer Metastasis to Kidney: An In Vitro Cell Line Based Study and Image Analysis

  • Aviral Kumar
  • Reetoja Nag
  • Satyakam Mishra
  • Bandaru Ramakrishna
  • V. V. R. Sai
  • Debasish MishraEmail author
Conference paper

Abstract

Metastasis of colorectal carcinoma to the kidney is a rare phenomenon and least-investigated mechanistically. Both exosomal and non-exosomal vesicles (NEVs) from tumor tissues have been proven to be important metastatic mediators. In this light, the current work focuses on the investigation of the role of NEVs obtained from colorectal cancer cell line HCT116 in developing metastatic traits in normal human embryonic kidney cell line HEK293. ECVs were isolated via filtration method from spent media of HCT116 culture. Dynamic light scattering (DLS) analysis showed ECVs which are obtained as a retentate of 220 nm filters had an average size of 147 nm and hence may be classified as non-exosomal vesicles. NEVs obtained from HCT116 spent media were poured onto compact culture plates of HEK293 cell lines. A systematic image analysis of crystal violet-stained plates was done using the snake model for segmentation by MATLAB and analysis by ImageJ. It is evident from the image analysis data that the number of disseminated cells/colony of cells was more in NEVs treated wells than that of the untreated ones. The average distance of centrifugal cell migration (analogous to invasion) was also found to be higher in case of nECV-treated HEK293 compact cultures. Although early, but in conclusion, it can be said that NEVs from colon carcinoma could be a metastatic mediator for human kidney cells. Secondly, it is indicated that 2D compact culture in combination with inexpensive image analytics can be a potential tool in anti-metaplastic drug discovery applications.

Keywords

Exosomes Non-exosomal vesicles Ultrafiltration Colorectal cancer Metastasis HCT116 Kidney HEK293 Snake-model image segmentation 

Notes

Acknowledgements

The authors acknowledge RGEMS seed fund, VIT University, Vellore for partially supporting the present research. The authors also would thank Dr. Everret R. Nelson for his timely support on cell lines.

Conflict of Interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Dulskas A, Bagurskas P, Sinkevicius Z, Samalavicius NE (2015) Sigmoid adenocarcinoma with metastases to the kidney: report of a rare case and review of the literature. Oncol Lett 10(2):1191–1193CrossRefGoogle Scholar
  2. 2.
    Suchorska WM, Lach MS (2016) The role of exosomes in tumor progression and metastasis (Review). Oncol Rep 35(3):1237–1244CrossRefGoogle Scholar
  3. 3.
    Pol VE, Bo AN (2012) Classification, functions, and clinical relevance of extracellular vesicles. Pharmacol Rev 64(3):676–705CrossRefGoogle Scholar
  4. 4.
    Soung YH, Nguyen T, Cao H, Lee J, Chung J (2015) Emerging roles of exosomes in cancer invasion and metastasis. BMB Rep 49(1):18–25CrossRefGoogle Scholar
  5. 5.
    Al-Nedawi K, Meehan B, Micallef J, Lhotak V, May L, Guha A, Rak J (2008) Intercellular transfer of the oncogenic receptor EGFRvIII by micro vesicles derived from tumor cells. Nat Cell Biol 10(5):619–624CrossRefGoogle Scholar
  6. 6.
    Lobb RJ, Becker M, Wen SW, Wong CSF, Wiegmans AP, Leimgruber A, Möller A (2015) Optimized exosome isolation protocol for cell culture supernatant and human plasma. J Extracellular Vesicles 4:27031CrossRefGoogle Scholar
  7. 7.
    Kramer N, Walzl A, Unger C, Rosner M, Krupitza G, Hengstschläger M, Dolznig H (2013) In vitro cell migration and invasion assays. Mutat Res 752(1):10–24CrossRefGoogle Scholar
  8. 8.
    Kumar KS, Pillong M, Kunze J, Burghardt I, Weller M, Grotzer MA, Schneider G, Baumgartner M (2015) Computer-assisted quantification of motile and invasive capabilities of cancer cells. Sci Rep 5:15338CrossRefGoogle Scholar
  9. 9.
    Gonzalez RC (2007) RE woods: digital image processing. Publishing House of Electronics Industry, pp 711–712Google Scholar
  10. 10.
    Amini AA, Weymouth TE, Jain TC (1990) Using dynamic programming for solving variational problems in vision. IEEE Trans Pattern Anal Mach Intell 12(9):855–867CrossRefGoogle Scholar
  11. 11.
    Kass M, Witkin A, Terzopoulos D (1988) Snake: active contour models. Int J Comput Vis 1(4):321–331CrossRefGoogle Scholar
  12. 12.
    Maletta F, Massa F, Torregrossa L, Duregon E, Casadei GP, Basolo F, Tallini G, Volante M, Nikiforov Y, Papotti M (2016) Cytological features of non-invasive follicular thyroid neoplasm with papillary-like nuclear features and their correlation with tumour histology. human pathology. Hum Pathol 54:134–142CrossRefGoogle Scholar
  13. 13.
    Bowen WR, Hilal N, Lovitt RW, William PM (1999) Atomic force microscope studies of membrane surfaces. In: TS Sorenson (ed) Surface chemistry and electrochemistry of membranes pp 1–38Google Scholar
  14. 14.
    Pol EV, Coumans FA, Grootemaat AE, Gardiner C, Sargent IL, Harrison P, Sturk A, van Leeuwen TG, Nieuwland R (2014) Particle size distribution of exosomes and microvesicles determined by transmission electron microscopy, flow cytometry, nanoparticle tracking analysis, and resistive pulse sensing. J Thromb Haemost 12(7):1182–1192CrossRefGoogle Scholar
  15. 15.
    Curl CL, Bellair CJ, Harris T, Allman BE, Harris PJ, Stewart AG, Roberts A, Nugent K, Delbridge LMD (2005) Refractive index measurement in viable cells using quantitative phase-amplitude microscopy and confocal microscopy. Cytometry 65A:88–92CrossRefGoogle Scholar
  16. 16.
    Oberemko AV, Popandopulo AG (2014) Extracellular vesicles: classification, functions and clinical relevance. Biotechnologia Acta 7(6):102–108CrossRefGoogle Scholar
  17. 17.
    Raposo G, Stoorvogel W (2013) Extracellular vesicles: Exosomes, microvesicles, and friends. J Cell Biol 200(4):373–383CrossRefGoogle Scholar
  18. 18.
    Minciacchi VR, Spinelli C, Reis-Sobreiro M, Cavallini L, You S, Zandian M, Li X, Mishra R, Chiarugi P, Adam RM, Posada EM, Viglietto G, Freeman MR, Cocucci E, Bhowmick NA, Di Vizio D (2017) MYC mediates large oncosome-induced fibroblast reprogramming in prostate cancer. Can Res 77(9):2306–2317CrossRefGoogle Scholar
  19. 19.
    Yoon YJ, Kim DK, Yoon CM, Park J, Kim YK, Roh TY, Gho YS (2014) Egr-1 activation by cancer-derived extracellular vesicles promotes endothelial cell migration via ERK1/2 and JNK signaling pathways. PLoS ONE 9(12):e115170CrossRefGoogle Scholar

Copyright information

© Springer Nature Singapore Pte Ltd. 2019

Authors and Affiliations

  1. 1.School of Bio-Sciences and Technology (SBST)Vellore Institute of TechnologyVelloreIndia
  2. 2.Centre for Biomaterials Cellular and Molecular Theranostics (CBCMT), Vellore Institute of TechnologyVelloreIndia
  3. 3.Department of Electronics and Communication EngineeringKattankulathur Campus, SRM Institute of Science and Technology (formerly known as SRM University)ChennaiIndia
  4. 4.Biomedical Engineering Laboratory, Department of Applied MechanicsIndian Institute of Technology MadrasChennaiIndia

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