Research Institute of Carcinogenesis

Moscow, Russia

Research Institute of Carcinogenesis

Moscow, Russia
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PubMed | Research Institute of Carcinogenesis, Moscow State University and RAS Engelhardt Institute of Molecular Biology
Type: | Journal: Scientific reports | Year: 2015

Exosomes, small (40-100nm) extracellular membranous vesicles, attract enormous research interest because they are carriers of disease markers and a prospective delivery system for therapeutic agents. Differential centrifugation, the prevalent method of exosome isolation, frequently produces dissimilar and improper results because of the faulty practice of using a common centrifugation protocol with different rotors. Moreover, as recommended by suppliers, adjusting the centrifugation duration according to rotor K-factors does not work for fixed-angle rotors. For both types of rotors--swinging bucket and fixed-angle--we express the theoretically expected proportion of pelleted vesicles of a given size and the cut-off size of completely sedimented vesicles as dependent on the centrifugation force and duration and the sedimentation path-lengths. The proper centrifugation conditions can be selected using relatively simple theoretical estimates of the cut-off sizes of vesicles. Experimental verification on exosomes isolated from HT29 cell culture supernatant confirmed the main theoretical statements. Measured by the nanoparticle tracking analysis (NTA) technique, the concentration and size distribution of the vesicles after centrifugation agree with those theoretically expected. To simplify this cut-off-size-based adjustment of centrifugation protocol for any rotor, we developed a web-calculator.


Botezatu I.V.,Research Institute of Carcinogenesis | Panchuk I.O.,Research Institute of Carcinogenesis | Stroganova A.M.,Research Institute of Carcinogenesis | Senderovich A.I.,Research Institute of Carcinogenesis | And 3 more authors.
Molecular Biology | Year: 2017

Scanning for mutations by DNA melting analysis (DMA) is based on asymmetric PCR followed by the melting of duplexes formed by single-stranded amplicons with TaqMan probes. The method is optimally suited for clinical genetic testing; it is easy to perform, high-throughput, and sensitive. The detection limit of mutant alleles by the DMA method is about 3%, which is much higher than the sensitivity of Sanger sequencing. In addition, the DMA method is realized in a closed-tube format, while 2-h assay is carried out in a single tube without any intermediate or additional procedures thereby minimizing the risk of cross contamination of the samples. The validation of the DMA method was performed by scanning for mutations of clinically significant genes KRAS, NRAS, BRAF, and PIK3CA in 324 DNA samples from tumors of patients with melanoma, colorectal and lung cancer. DNA was isolated either directly from tumor tissues, or from formalin- fixed paraffin-embedded tumor tissues. The detected mutations were verified by Sanger sequencing. The spectra of mutations identified in each tumor type correspond to the literature data and, thus, validate the use of DMA. © 2017, Pleiades Publishing, Inc.


Kovaleva O.V.,Research Institute of Carcinogenesis | Efremov G.D.,National Medical Research Radiological Center | Mikhaylenko D.S.,National Medical Research Radiological Center | Alekseev B.Y.,National Medical Research Radiological Center | Grachev A.N.,Research Institute of Carcinogenesis
Onkourologiya | Year: 2017

The role of tumor stroma in malignant tumor pathogenesis cannot be disputed. Macrophages are one of the crucial elements of tumor stroma. Tumor-associated macrophages (TAMs) are type 2-activated macrophages (M2). They were first described in 1992. They carry CD206, CD163, FXIIIa, βIG-H3, stabilin 1, YKL-39, SI-CLP, tenascin C, LOX-1, fibronectin, MARCO, interleukin 1 receptor antagonist (IL-1RA) and other markers. Unlike proinflammatory macrophages (M1), M2 display high anti-inflammatory activity and are responsible for inflammation reaction suppression and tissue recovery in inflamed area. TAMs significantly contribute to tumor progression by stimulating cell proliferation, angiogenesis, and suppression of antitumor immune response. Identification of macrophages in renal tumors involves a limited number of markers, which doesn't allow making a conclusive answer about their function. However, a correlation between TAMs content and a negative disease prognosis can be considered proven. Studies of M1 and M2 using different markers have shown that renal tumors contain high levels of TAMs with mixed M1/M2 phenotype. TAMs in renal tumors are highly proangiogenic and immunosuppressive. TAMs density can be used as a prognostic marker, but development of an effective treatment strategy aimed at inhibition of TAMs antitumor activity requires systemic research involving a wide panel of M1 and M2 macrophage markers.


Kalitin N.N.,Research Institute of Carcinogenesis
Bulletin of experimental biology and medicine | Year: 2012

We studied the expression of genes encoding vascular endothelial growth factors VEGF-A, VEGF-C, VEGF-D and their receptors in cell cultures of human multiple myeloma IM9, RPMI 1640, RPMI 8226. The studied cells did not differ by the expression of growth factors. Expression of VEGFR1 receptor was detected only in IM9 cells and VEGFR2 and VEGFR3 receptors were not expressed in multiple myeloma cells. A dependence between the aberrant CD45/CD56 phenotype of human multiple myeloma cells and VEGFR1 expression in them was revealed. The only VEGFR1-positive IM9 cell culture was most resistant to Velcade (bortezomib).


PubMed | Research Institute of Carcinogenesis
Type: Journal Article | Journal: Bulletin of experimental biology and medicine | Year: 2012

We studied the expression of genes encoding vascular endothelial growth factors VEGF-A, VEGF-C, VEGF-D and their receptors in cell cultures of human multiple myeloma IM9, RPMI 1640, RPMI 8226. The studied cells did not differ by the expression of growth factors. Expression of VEGFR1 receptor was detected only in IM9 cells and VEGFR2 and VEGFR3 receptors were not expressed in multiple myeloma cells. A dependence between the aberrant CD45/CD56 phenotype of human multiple myeloma cells and VEGFR1 expression in them was revealed. The only VEGFR1-positive IM9 cell culture was most resistant to Velcade (bortezomib).


Morozova O.V.,Research Institute of Carcinogenesis | Karamysheva A.E.,Research Institute of Carcinogenesis | Shavochkina D.A.,Research Institute of Carcinogenesis
Voprosy Onkologii | Year: 2013

There was examined the role of VEGFA in the induction of malignant hemangioendothelioma (HAE) in the nonvascular tissue of renal capsule (RC) in mice. Expression mRNA VEGFA and its receptor Flk1 in RC and the kidneys in male mice of CBA and C57B1/6 lines was studied at different stages of the administration of a chemical carcinogen 1,2-dimethylhydrazine (DMH), which induced HAE of PC in CBA males with a high frequency, and in C57B1/6 line - with a low frequency. In the kidneys of male mice of both lines at all stages there was observed significant expression both VEGFA and Flkl but no linear differences were found. In RC of these mice expression of studied genes were not found. Only at the last stage of carcinogenesis in RC of resistant line C57B1/6 and in HAE of RC of line CBA there was appeared weak expression of mRNA VEGFA but mRNA Flk1 was not expressed. A similar profile of expression is appropriate more for pericytes and macrophages but not endothelial cells indicating either on the origin of HAE RC from non-endothelial cells or on atypia of endothelial cells - progenitors of this tumor.

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