The obtaining and characterization new soft tissue sarcoma and osteogenic sarcoma cell lines for translational research



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Abstract

The rare occurrence, high histological heterogeneity, changing classification standards, and complexity of cultivation are the reasons for the failure of soft tissue and bone sarcomas (STBS) cell lines required for large-scale preclinical studies. Obtaining and characterization new STBS cell lines is the great importance for creating cell models that allow us to study the processes of oncogenesis and metastasis of mesenchymal tumors in the development of new treatment methods. Purpose: to create a collection of new characterized STBS cell lines suitable for translational research. In the study were used tumor samples from 71 patients of the N.N. Petrov National Medical Research Center of Oncology with the diagnosis of "sarcoma”. The samples were received during the operation in period 2013-2020. A standard protocol was developed for the cultivation of STBS cells. The proliferative, invasive and migration activity of the obtained cell lines in monolayer and spheroids was studied, HLA typing was performed, and the expression of cancer-testicular genes (CTG) and stem cell markers (CD133 and ALDH1). A collection of STBS cell lines, which includes 54 stable cell lines of 18 histological subtypes, including 39 soft tissue and 15 bone sarcomas cultures, with the metastatic culture of 81.5% was created (n=44). In the process of long-term cultivation there was an increase in the proliferative activity of sarcomas cells (p<0,05). Statistically significant differences in migration activity parameters between soft tissue and bone sarcomas cultures were revealed (p<0,05). A high degree of heterogeneity of the transcriptional activity of the studied CTG was found, and the absence of expression was detected in 19.2% of cases. The activity of the PRAME (55.8%) and GAGE1 (50%) genes was detected with the highest frequency (rho=0.5025; p=0.00015). PASD1 expression correlated with GAGE1 (rho=0.6951; p=0.00001), PRAME (rho=0.5743; p=0.00001). Co-expression of the NY-ESO-1 and MAGEA1 genes was also detected: (rho=0.4027; p=0.00308). The inverse correlation of average strength between the number of ALDH1 + cells and CD133+cells in cultures and the value of time to progression in patients (rho=-0.505, p=0.033; rho=-0.513, p=0.021, respectively) was established. The received and characterized cell lines can become an important component for realization of perspective translational research, a demanded tool for solution of various tasks of modern medicine, such as proteomic research, revealing of important signal mechanisms, search of target molecules for creation of new therapeutic approaches.

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About the authors

N. A Avdonkina

N.N. Petrov National Medical Research Center of Oncology

Email: nataliaavdonkina@gmail.com

A. B Danilova

N.N. Petrov National Medical Research Center of Oncology

V. A Misyurin

N.N. Blokhin National Medical Research Center of Oncology

E. A Prosekina

N.N. Petrov National Medical Research Center of Oncology

N. V Emelyanova

N.N. Petrov National Medical Research Center of Oncology

T. L Nekhaeva

N.N. Petrov National Medical Research Center of Oncology

O. V Skachkova

N.N. Petrov National Medical Research Center of Oncology

A. V Novik

N.N. Petrov National Medical Research Center of Oncology

N. P Pipia

N.N. Petrov National Medical Research Center of Oncology

G. I Gafton

N.N. Petrov National Medical Research Center of Oncology

E. V Levchenko

N.N. Petrov National Medical Research Center of Oncology

A. M Belyaev

N.N. Petrov National Medical Research Center of Oncology

I. A Baldueva

N.N. Petrov National Medical Research Center of Oncology

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