Abstract

Background. Heat stress (HS) induces the cellular secretion of heat shock proteins (HSP ) and extracellular nanovesicles (ENVs). The biological link between these phenomena is poorly understood. In the case of colorectal cancer (CRC) cells, the secretion of HSP s and ENV may be involved in the clinical response to intraperitoneal therapy of peritoneal carcinomatosis.Material and Methods. Established colon cancer cell lines COLO 320, HCT 116, HT29 and DLD 1 were used. ENVs were isolated from culture media by differential ultra-centrifugation and analyzed by dynamic light scattering, nanoparticle tracking analysis, atomic force microscopy and flow cytometry. Super-paramagnetic particles (SPMP ) covered by antibodies to the membrane form of Hsp70 were used for isolation and quantification of Hsp70(+) ENVs. Vesicular microRNA was assayed by RT-qPC R.Results. HS induces the secretion of ENVs by CRC cells, the resistance to HS correlates with the activity of HS-induced ENVs secretion. HS induces the secretion of a specific population of ENVs enriched by membrane form Hsp70 (mHsp70). The microRNA content of mHsp70(+) ENVs has qualitative and quantitative features. The concentration of miR-126-3p, -181-5p, -155-5p, -223 is increased in mHSP 70(+) ENVs secreted by three CRC cell lines.Conclusion. HS induces the secretion of mHSP 70(+) ENVs by CRC cells. This phenomenon may be involved in a clinical response to intraperitoneal chemo-hyperthermic perfusion therapy of peritoneal carcinomatosis.

Highlights

  • Heat stress (HS) induces the cellular secretion of heat shock proteins (HSP) and extracellular nanovesicles (ENVs)

  • In the case of colorectal cancer (CRC) cells, the secretion of HSPs and ENV may be involved in the clinical response to intraperitoneal therapy of peritoneal carcinomatosis

  • Super-paramagnetic particles (SPMP) covered by antibodies to the membrane form of Hsp70 were used for isolation and quantification of Hsp70(+) ENVs

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Summary

Laboratory and experImental studies

I.V. Nazarova1, L.M. Zabegina1, N.S. Nikiforova1, M.A. Slusarenko1, E.I. Sidina1, A.V. Zhakhov3, A.M. Ishchenko3, B.A. Margulis4, I.V. Guzhova4, A.V. Malek1,2 N.N. Petrov National Medical Research Center of Oncology, Saint Petersburg, Russia1 68, Leningradskaya St., 197758, Pesochny, Saint Petersburg, Russia1 Oncosystem Ltd., Moscow, Russia2 7, Nobel St., 7121205, Skolkovo Innovation Center, Moscow, Russia2 Institute of Highly Pure Biopreparations, Saint Petersburg, Russia3 7, Pudozhsakya St., 197110, Saint Petersburg, Russia3 Institute of Cytology, RAS, Saint Petersburg, Russia4 4, Tikhoretsky Ave., 194064, Saint Petersburg, Russia4

Effect of HS on ENVs secretion activity assessed by NTA
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