Radiation effects on early phase of NT2/D1 neural differentiation in vitro

Abstract

Purpose: Widespread medical use of radiation in diagnosis, imaging and treatment of different central nervous system malignancies lead to various consequences. Aim of this study was to further elucidate mechanism of cell response to radiation and possible consequence on neural differentiation.

Materials and methods: NT2/D1 cells that resemble neural progenitors were used as a model system. Undifferentiated NT2/D1 cells and NT2/D1 cells in the early phase of neural differentiation were irradiated with low (0.2 Gy) and moderate (2 Gy) doses of γ radiation. The effect was analyzed on apoptosis, cell cycle, senescence, spheroid formation and the expression of genes and miRNAs involved in the regulation of pluripotency or neural differentiation.

Results: Two grays of irradiation induced apoptosis, senescence and cell cycle arrest of NT2/D1 cells, accompanied with altered expression of several genes (SOX2, OCT4, SOX3, PAX6) and miRNAs (miR-219, miR-21, miR124-a). Presented results show that 2 Gy of radiation significantly affected early phase of neural differentiation in vitro.

Conclusions: These results suggest that 2 Gy of radiation significantly affected early phase of neural differentiation and affect the population of neural progenitors. These findings might help in better understanding of side effects of radiotherapy in treatments of central nervous system malignancies.

Keywords:

• NT2/D1 cell line
• radiation
• neural differentiation
• miRNAs
• markers of pluripotency

Disclosure statement

The authors report no conflicts of interest.

Additional information

Funding

This work was supported by the Ministry of Education, Science and Technological Development, Republic of Serbia (Grant No 173051), the Serbian Academy of Sciences and Arts (Grant No: F 24), IBRO/PERC InEurope Short Stay Grants and EURATOM Fission and European Commission 7^th Framework Programme, DarkRisk (contract number 323216).

Notes on contributors

Danijela Stanisavljevic

Dr. Danijela Stanisavljevic is a Research Associate in Human Molecular Genetics Laboratory, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade. Her research interest: molecular mechanisms involved in neural development and malignant transformation of cell.

Jelena Popovic

Dr. Jelena Popovic is a Senior Research Associate in Human Molecular Genetics Laboratory, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade and a member of the Scientific Council of the same Institution. Her research interests: molecular pathways involved in cancer development/progression and improving efficiency in radiation therapy.

Isidora Petrovic

Dr. Isidora Petrovic is a Research Associate in Human Molecular Genetics Laboratory, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade. Her research interest: human molecular genetics, cancer biology, screening for novel bioactive and synthetic drugs.

Slobodan Davidovic

Dr. Slobodan Davidovic is a Research Associate in Human Molecular Genetics Laboratory, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade. His research interests: analysis of molecular diversity of Serbian population based on mitochondrial DNA polymorphisms and analysis of cytotoxic and anti-cancerogenic potential of bioactive compounds isolated from fungus/plants.

Michael J. Atkinson

Prof. Dr. Michael J. Atkinson is a full Professor at Medical Faculty, Technical University Munich (TUM) and Director of the Institute of Radiation Biology at Helmholtz Zentrum München, with research interest: radiation biology, radiation protection, molecular genetics.

Nataša Anastasov

Dr. Nataša Anastasov is a principal investigator at Institute of Radiation Biology, Helmholtz Zentrum München and lecturer at Medical Faculty, Technical University Munich (TUM) for program Master in Radiation Biology. Her research interest: radiation biology and oncology, drug screening, lentiviral gene therapy tools.

Milena Stevanovic

Academician Milena Stevanovic is full member of Serbian Academy of Sciences and Arts, full Professor at Faculty of Biology, University of Belgrade, Head of Human Molecular Genetics Laboratory, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade. Research interest: human molecular genetics, maintaining pluripotency and neural differentiation of stem cells; cytotoxic, antioxidant and anticancer properties of natural and synthetic compounds, molecular phylogenetic.