Translartion. Region: Russians Fedetion –
Source: Novosibirsk State University – Novosibirsk State University –
Scientists from Novosibirsk State University, together with colleagues from the Research Institute of Clinical and Experimental Lymphology (NIIKEL, a branch of the Institute of Cytology and Genetics SB RAS, ICG SB RAS), the Institute of Cytology and Genetics SB RAS (ICG SB RAS) and the G. I. Budker Institute of Nuclear Physics (INP SB RAS) have established that terahertz radiation affects the energy metabolism of melanoma cells. To this end, they conducted experiments to study the effects of this type of electromagnetic radiation on human melanoma cells. This work is of a fundamental nature and expands our understanding of the biological effects of terahertz radiation, as well as cellular reactions to its effects. The results are published in the journal “Biochimica et Biophysica Acta (BBA) – Molecular and Cell Biology of Lipids”.
Terahertz radiation (THzI) is electromagnetic waves whose frequency lies between the infrared and ultra-high-frequency (UHF) ranges: from 100 GHz to 10 THz. Modern technologies based on the use of electromagnetic waves in the terahertz range are widely used in biomedical sciences. For example, terahertz spectroscopy can be relevant in medical practice for the diagnosis of oncological diseases. At the same time, the THz region has not been fully studied, so fundamental research into the radiation of this electromagnetic spectrum and, first of all, the study of its biological effects on living systems are relevant.
— Our work is devoted to studying the fundamental mechanisms of the impact of non-ionizing radiation on biological objects, in this case, on human melanoma cells. However, the purpose of the study is not to develop treatment methods using terahertz radiation. We chose a melanoma cell line as a model, since it is a stable and well-studied system. This allows us to minimize the impact of side factors and be sure that the observed changes are associated with the impact of THz radiation, and not with the features of the cells’ vital activity, — the first-year postgraduate student commented Faculty of Natural Sciences of NSU (major in biology), junior researcher at the laboratory of cell technologies at the Research Institute of Cellular and Electron Microbiology and Genetics, a branch of the Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences, Ekaterina Butikova.
These studies were conducted at the Novosibirsk Free Electron Laser (NFEL) of the Institute of Nuclear Physics SB RAS. Only this facility can generate radiation with the parameters required for these experiments: the frequency of the radiation used was 2.3 THz, and the average intensity was 0.05 W/cm2. The specialists exposed human melanoma cells grown in culture flasks to THzI. Irradiation at a radiation frequency of 2.3 THz was carried out at the user station of the Novosibirsk Free Electron Laser.
– The Novosibirsk LSE is a unique source of teragerz and infrared radiation. In terms of average power, it is many orders of magnitude exceeds any sources existing in the world, which allows you to conduct absolutely unique experiments in a very wide area of wavelengths with various biological objects. The fact is that biopolymers, such as proteins, have four spatial levels of organization. If the primary structure is determined by covalent bonds, then the secondary, tertiary and higher are determined by hydrogen bonds, the energy of which lies precisely in the area of TGC-radiation. Therefore, if we affect the TGCI on living systems, we can quite much affect the operation of their cells, on the processes that pass inside them. Such experiments are of interest from the point of view that no living organism has formed any protective mechanisms from TGC radiation, since it is completely absorbed by the atmosphere, which means that it affects the biological objects, it can be explored how they adapt, which protection mechanisms include. For such biological experiments, a special user station was created on NLSE, which implemented the technology for adjusting the average and peak radiation power, as well as the intensity of exposure. Since we work with living systems that feel comfortable in a very narrow temperature range, which was important for the purity of experiments to equip the station with a wiper and thermal imager – these devices support and control the desired temperature. Thanks to this, we understand that we get the reaction of the system precisely to the influence of irradiation, and not to the increase or decrease in temperature, ”explained Vasily Popik, senior researcher at the Physical and Mathematical Sciences of the Physical and Mathematics.
Three groups of cells participated in the experiment. One was irradiated with terahertz radiation, the second with infrared radiation (IR), and the third was a control group and was not affected in any way. The terahertz and IR groups were irradiated for 10 and 45 minutes. On the day of irradiation, specialists conducted cytotoxic tests on the cells. On the third day, they conducted metabolomic screening – an analysis of metabolites, or organic molecules involved in metabolism.
– Metabolites are small organic molecules that are involved in the metabolism in living organisms. They can be intermediate or final products of biochemical reactions, provide cells with energy, serve as a building material for cells or perform regulatory functions. In the course of complex biochemical transformations, some substances are synthesized, others are destroyed, ensuring the energy balance, biosynthesis and the regulation of cellular functions. To study the biochemical state of cells and tissues, one of the most effective tools is metabolo screening. It allows you to fix changes in the metabolic composition of the body associated with physiological processes, diseases or external influences. Analysis of a wide range of metabolites helps to look into the molecular world of the cell and understand how it functions. In our laboratory, we conduct metabolon screening by the method of highly effective liquid chromatography with tandem mass-spectrometric detection (VEZH-MS/MS). Two years ago, we developed an approach that allows you to analyze about 400 metabolites (including both polar compounds and lipids) in less than 30 minutes of analysis. This was made possible thanks to the use of a monolithic column for VEGH, created by the employees of the Catalysis Institute SB RAS Yu.S. Sotnikova and Yu.V. Patrushev, ”said the laboratory assistant of the laboratory of the molecular pathology of the Institute of Medicine and Medical Technology of NSU, junior researcher at the Laboratory of Physiologically active substances of the Novosibirsk Institute of Organic Chemistry named after N.N. Vorozhtsova SB RAS (Nioh SB RAS) Nikita Basov.
The scientists have previously applied their metabolomic screening approach to plasma and dried blood spots, but its use in cell culture studies remained unexplored. In this work, they developed and tested a cell sample preparation protocol, assessed its limitations, and combined it for the first time with an analytical method to study the effects of terahertz radiation on melanoma cells.
Using metabolomic screening data and bioinformatics tools, the team of scientists concluded that terahertz radiation primarily affects the cell’s energy metabolism. To do this, they used the ANDSystem tool, an automated system that combines data from numerous biological databases and scientific publications, allowing them to identify functional links between genes, proteins, and metabolic pathways.
— Our studies show that THz radiation caused changes in the content of 40 metabolites, mainly in the pathways of purine and pyrimidine metabolism, and it also affects the level of ceramides and phosphatidylcholines. Analysis of genetic networks conducted by our colleagues from the Laboratory of Computer Proteomics of the Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences identified mitochondrial membrane proteins as key regulators of the biosynthesis of these metabolites. In addition, THz radiation apparently disrupts the structure of lipid rafts, which affects mitochondrial transport, but does not affect the integrity of proteins. Metabolic effects were specific to THzI and differed from the thermal effects observed with infrared radiation, — added Ekaterina Butikova.
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