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Title: The characterization of exosome from blood plasma of patients with colorectal cancer

Abstract

Exosomes are extracellular membrane structures involved in many physiological and pathological processes including cancerogenesis and metastasis. The clarification of the criteria for exosome isolating and identifying is the purpose of this study. Exosome samples from the plasma of patients with colorectal cancer and healthy donors were examined using transmission electron microscopy and flow cytometry in accordance with the minimum requirements of “International Society for Extracellular Vesicles”. The choice of the method for isolation of exosomes from the blood plasma by ultrafiltration and ultracentrifugation allowed obtaining highly purified samples of exosomes, in which all the structural components were clearly seen. The results obtained with flow cytometry suggest that exosomes of blood plasma from patients with colorectal cancer can be produced by epithelial cells. Moreover, cells produce different types of exosomes, which correspond to different mechanisms in sorting macromolecules in the membrane of multivesicular bodies. Determination of significant differences in the expression of specific exosomal proteins from colorectal cancer patients compared to healthy donors suggests a high diagnostic potential significance of circulating exosomes.

Authors:
 [1];  [2];  [3];  [2]; ; ;  [1];  [4]
  1. Tomsk Cancer Research Institute, Kooperativny Street 5, Tomsk, 634009 (Russian Federation)
  2. (Russian Federation)
  3. Institute of Chemical Biology and Fundamental Medicine SB RAS, Lavrentiev Avenue 8, Novosibirsk, 630090 (Russian Federation)
  4. Siberian State Medical University, Moskovsky Trakt 2, Tomsk, 634050 (Russian Federation)
Publication Date:
OSTI Identifier:
22608294
Resource Type:
Journal Article
Resource Relation:
Journal Name: AIP Conference Proceedings; Journal Volume: 1760; Journal Issue: 1; Conference: PC'16: International conference on physics of cancer: Interdisciplinary problems and clinical applications 2016, Tomsk (Russian Federation), 22-25 Mar 2016; Other Information: (c) 2016 Author(s); Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE; BLOOD PLASMA; COMPARATIVE EVALUATIONS; ELECTRONS; MEMBRANES; METASTASES; NEOPLASMS; PATIENTS; PLASMA; TRANSMISSION; TRANSMISSION ELECTRON MICROSCOPY; ULTRACENTRIFUGATION; ULTRAFILTRATION

Citation Formats

Yunusova, N. V., E-mail: Bochkarevanv@oncology.tomsk.ru, Siberian State Medical University, Moskovsky Trakt 2, Tomsk, 634050, Tamkovich, S. N., E-mail: s.tamk@niboch.nsc.ru, Novosibirsk State University, Pirogov Street 2, Novosibirsk, 630090, Stakheeva, M. N., E-mail: StakheyevaM@oncology.tomsk.ru, Afanas’ev, S. G., E-mail: Afanasievsg@oncology.tomsk.ru, Kondakova, I. V., E-mail: Kondakova@oncology.tomsk.ru, and Frolova, A. Y., E-mail: Frolovalenya@mail.ru. The characterization of exosome from blood plasma of patients with colorectal cancer. United States: N. p., 2016. Web. doi:10.1063/1.4960289.
Yunusova, N. V., E-mail: Bochkarevanv@oncology.tomsk.ru, Siberian State Medical University, Moskovsky Trakt 2, Tomsk, 634050, Tamkovich, S. N., E-mail: s.tamk@niboch.nsc.ru, Novosibirsk State University, Pirogov Street 2, Novosibirsk, 630090, Stakheeva, M. N., E-mail: StakheyevaM@oncology.tomsk.ru, Afanas’ev, S. G., E-mail: Afanasievsg@oncology.tomsk.ru, Kondakova, I. V., E-mail: Kondakova@oncology.tomsk.ru, & Frolova, A. Y., E-mail: Frolovalenya@mail.ru. The characterization of exosome from blood plasma of patients with colorectal cancer. United States. doi:10.1063/1.4960289.
Yunusova, N. V., E-mail: Bochkarevanv@oncology.tomsk.ru, Siberian State Medical University, Moskovsky Trakt 2, Tomsk, 634050, Tamkovich, S. N., E-mail: s.tamk@niboch.nsc.ru, Novosibirsk State University, Pirogov Street 2, Novosibirsk, 630090, Stakheeva, M. N., E-mail: StakheyevaM@oncology.tomsk.ru, Afanas’ev, S. G., E-mail: Afanasievsg@oncology.tomsk.ru, Kondakova, I. V., E-mail: Kondakova@oncology.tomsk.ru, and Frolova, A. Y., E-mail: Frolovalenya@mail.ru. 2016. "The characterization of exosome from blood plasma of patients with colorectal cancer". United States. doi:10.1063/1.4960289.
@article{osti_22608294,
title = {The characterization of exosome from blood plasma of patients with colorectal cancer},
author = {Yunusova, N. V., E-mail: Bochkarevanv@oncology.tomsk.ru and Siberian State Medical University, Moskovsky Trakt 2, Tomsk, 634050 and Tamkovich, S. N., E-mail: s.tamk@niboch.nsc.ru and Novosibirsk State University, Pirogov Street 2, Novosibirsk, 630090 and Stakheeva, M. N., E-mail: StakheyevaM@oncology.tomsk.ru and Afanas’ev, S. G., E-mail: Afanasievsg@oncology.tomsk.ru and Kondakova, I. V., E-mail: Kondakova@oncology.tomsk.ru and Frolova, A. Y., E-mail: Frolovalenya@mail.ru},
abstractNote = {Exosomes are extracellular membrane structures involved in many physiological and pathological processes including cancerogenesis and metastasis. The clarification of the criteria for exosome isolating and identifying is the purpose of this study. Exosome samples from the plasma of patients with colorectal cancer and healthy donors were examined using transmission electron microscopy and flow cytometry in accordance with the minimum requirements of “International Society for Extracellular Vesicles”. The choice of the method for isolation of exosomes from the blood plasma by ultrafiltration and ultracentrifugation allowed obtaining highly purified samples of exosomes, in which all the structural components were clearly seen. The results obtained with flow cytometry suggest that exosomes of blood plasma from patients with colorectal cancer can be produced by epithelial cells. Moreover, cells produce different types of exosomes, which correspond to different mechanisms in sorting macromolecules in the membrane of multivesicular bodies. Determination of significant differences in the expression of specific exosomal proteins from colorectal cancer patients compared to healthy donors suggests a high diagnostic potential significance of circulating exosomes.},
doi = {10.1063/1.4960289},
journal = {AIP Conference Proceedings},
number = 1,
volume = 1760,
place = {United States},
year = 2016,
month = 8
}
  • Purpose. We retrospectively evaluated the safety and efficacy of preoperative initial hepatic arterial infusion chemotherapy (HAIC) through a port-catheter system in patients with liver dysfunction due to synchronous and unresectable liver metastases. The aim of HAIC was to improve patients' clinical condition for later surgical removal of primary colorectal cancer. Methods. Port-catheter systems were placed radiologically in 21 patients (mean age 58.6 {+-} 8.1 years) with liver dysfunction due to synchronous liver metastases from colorectal cancer. Initial HAIC of 1,000 mg/m{sup 2} 5-fluorouracil was administered weekly as a 5 hr continuous infusion through this system. Surgical removal of the primarymore » lesion was planned after HAIC improved the liver function. Results. Port-catheter system placement was successful in all patients without severe complications. Patients were followed up for a median of 309 days (range 51-998 days). After starting HAIC, no severe adverse events that caused drug loss and treatment postponement or suspension were observed in any of the patients. HAIC was performed a mean of 4.5 {+-} 3.0 times and the liver function improved in all patients. Curative (n = 18) or palliative (n = 1) surgical removal of the primary lesion was performed. The remaining 2 patients died because extrahepatic metastases developed and their performance status worsened; thus, surgery could not be performed. The median survival times of all patients and the operated patients were 309 and 386 days, respectively. Conclusion. Initial HAIC administration is a safe and efficacious method for improving liver function prior to operative resection of primary colorectal cancer in patients with liver dysfunction due to synchronous and unresectable liver metastases.« less
  • The purpose of this study was to assess retrospectively the sequential treatment of hepatic arterial infusion (HAI) chemotherapy followed by systemic therapy using oxaliplatin plus 5-flourouracil (5-FU) and leucovorin, namely, FOLFOX, for patients with liver metastases from colorectal cancer. We reviewed 20 patients with unresectable liver metastases from colorectal cancer. Patients were initially treated with HAI chemotherapy until disease progression (5-fluorouracil, 1000 mg/m{sup 2} intra-arterial infusion, weekly) and then with FOLFOX thereafter (FOLFOX4, n = 13; modified FOLFOX6, n = 7). Adverse events, tumor response, and time to progression for each therapy were evaluated retrospectively, and overall survival was estimated.more » Toxicity of HAI chemotherapy was generally mild. Of 20 patients, adverse events leading to treatment discontinuation occurred in only 1 patient (5%) during initial therapy using HAI chemotherapy, while 9 patients (45%) exhibited adverse events during subsequent FOLFOX therapy. For HAI chemotherapy and FOLFOX, objective response rates were 85.0% and 35.0%, respectively, and median time to progression was 11.6 and 5.1 months, respectively. Median overall survival was 30.1 months. In conclusion, the sequence of HAI chemotherapy followed by FOLFOX is a promising treatment strategy for the long-term use of active chemotherapeutic agents, leading to a superior tumor response and fewer toxic effects in patients with unresectable liver metastases from colorectal cancer.« less
  • Purpose: Predictive assays for acute radiation toxicities would be clinically relevant in radiation oncology. We prospectively examined the predictive role of the survival fraction at 2 Gy (SF2) and of γH2AX (double-strand break [DSB] DNA marker) expression kinetics in peripheral blood mononuclear cells (PBMCs) from cancer patients before radiation therapy. Methods and Materials: SF2 was measured with Trypan Blue assay in the PBMCs from 89 cancer patients undergoing radiation therapy at 4 hours (SF2{sub [4h]}) and 24 hours (SF2{sub [24h]}) after ex vivo irradiation. Using Western blot analysis and band densitometry, we further assessed the expression of γH2AX in PBMC DNA at 0 hours, 30 minutes,more » and 4 hours (33 patients) and 0 hour, 4 hours, and 24 hours (56 patients), following ex vivo irradiation with 2 Gy. Appropriate ratios were used to characterize each patient, and these were retrospectively correlated with early radiation therapy toxicity grade. Results: The SF2{sub (4h)} was inversely correlated with the toxicity grade (P=.006). The γH2AX-ratio{sub (30min)} (band density of irradiated/non-irradiated cells at 30 minutes) revealed, similarly, a significant inverse association (P=.0001). The DSB DNA repair rate from 30 minutes to 4 hours, calculated as the relative RγH2AX-ratio (γH2AX-ratio{sub (4h)}/γH2AX-ratio{sub (30min)}) showed a significant direct association with high toxicity grade (P=.01). Conclusions: Our results suggest that SF2 is a significant radiation sensitivity index for patients undergoing radiation therapy. γH2AX Western blot densitometry analysis provided 2 important markers of normal tissue radiation sensitivity. Low γH2AX expression at 30 minutes was linked with high toxicity grade, suggesting that poor γH2AX repair activity within a time frame of 30 minutes after irradiation predicts for poor radiation tolerance. On the other hand, rapid γH2AX content restoration at 4 hours after irradiation, compatible with efficient DSB repair ability, predicts for increased radiation tolerance.« less
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