Karyotype alterations in human lung adenocarcinoma cells after long-term action of interferon-alpha

Aim: To estimate the effect of long-term IFN treatment of human non-small-cell lung cancer cell line A-549 on their karyotype characteristics and on the clonal structure of cell population. Methods: Cytogenetic research was performed by standard methods using routine and differential staining. Cytogenetic characteristics were estimated per 1000 cells (ppm, (‰)). Results: Cytogenetic analysis of IFN-modified A-549 human lung cancer cells had demonstrated far-going changes in their population structure. It was shown that long term cultivation with IFN altered the chromosome modal class of A-549 cells, induced the domination of hromosomes with certain molecular markers: the number of metaphases with der (6) t (6; 1) chromosomal rearrangement increased significantly (from 6% to 80%, p < 0.001) and the cells with der (2) t (2; 1) markers almost disappeared. Thus, under the effect of IFN the cell clonal selection takes place. Decrease of the cell division rate and pseudometaphase occurrence, increase of the number of cells containing micronuclei are the typical characteristics of IFN-modified А-549 cell subline A-549IFN. Conclusion: Long-term IFN effect results in alterations of cytogenetic properties of A-549 human lung cancer cells

Spontaneous premature condensation of chromosomes in normal and transformed mammal cells

Aim: To study the relation between premature chromosome condensation and the ability of the cells to undergo malignant transformation. Methods: Standard cytogenetic analysis of bone marrow cells and cultured normal and tumor cells has been used. Results: Comparative analysis of the frequency of occurrence of the cells with premature chromosome condensation (РСС) (cell “arrest” at G2/M phase) in relation to dividing cells in the cultures of human immortalized cells of hematopoietic origin, human lung carcinoma A-549 cells, and in populations of bone marrow cells of BALB/c and C57BL/6 mice differing in predisposition for myeloma development has been performed. It has been revealed that in populations of bone marrow cells of C57BL/6 mice the relation of cells with РСС to dividing ones is 2–3-fold lower than in other studied cell populations. Immortalized and malignantly transformed human cell lines were characterized by high frequency of occurrence of cells with РСС. In the cells of А-549R subline characterized by suppressed malignant phenotype this index was lower than in parental А-549 cells. Conclusion: The obtained data point on possible relation between disturbed passing of “check point” by cells upon transition from G2 phase of cell cycle to mitosis and increased genetic heterogeneity of new cell generation associated with ability of cells to immortalization and malignant transformation.Цель: изучение взаимосвязи между преждевременной конденсацией хромосом и склонностью клеток к злокачественной трансформации. Методы: стандартные методы цитогенетического анализа клеток костного мозга и культивируемых нормальных и опухолевых клеток. Результаты: проведен сравнительный анализ отношения частоты выявления клеток с преждевременной конденсацией хромосом (РСС) (арест клеток в G2/M) к делящимся клеткам в культурах иммортализованных клеток человека кроветворного происхождения и злокачественно трансформированных клеток рака легкого человека линии A-549, а также в популяциях клеток костного мозга двух линий мышей: линии BALB/c с высокой предрасположенностью к развитию миеломы и C57BL/6, без такой предрасположенности. Выявлено, что в популяциях клеток костного мозга мышей низкораковой линии C57BL/6 отношение клеток с РСС к делящимся в 2–3 раза меньше, чем в других исследованных клеточных популяциях. Иммортализованные и злокачественно трансформированные клеточные линии человека характеризовались высокой частотой присутствия клеток с РСС. В клетках сублинии А-549R, характеризующейся подавлением признаков злокачественности, этот показатель заметно ниже, чем в клетках исходной линии А-549. Выводы: полученные данные позволяют предположить связь между нарушением прохождения клетками точки проверки при переходе от G2 фазы клеточного цикла к митозу и повышенной генетической гетерогенностью их потомства, ассоциированной со склонностью клеток к иммортализации и злокачественной трансформации

Resource-saving technology for oily wastewater treatment in agro-industrial complexes

The authors of the article consider the problem of protection of water bodies from pollution by oily wastewater from the territory of enterprises of the agro-industrial complex engineering and technical systems. A resource-saving technology for this wastewater treatment using multicomponent sorbents has been developed

Genetic diversity of SAD and FAD genes responsible for the fatty acid composition in flax cultivars and lines

Background: Flax (Linum usitatissimum L.) is grown for fiber and seed in many countries. Flax cultivars differ in the oil composition and, depending on the ratio of fatty acids, are used in pharmaceutical, food, or paint industries. It is known that genes of SAD (stearoyl-ACP desaturase) and FAD (fatty acid desaturase) families play a key role in the synthesis of fatty acids, and some alleles of these genes are associated with a certain composition of flax oil. However, data on genetic polymorphism of these genes are still insufficient. Results: On the basis of the collection of the Institute for Flax (Torzhok, Russia), we formed a representative set of 84 cultivars and lines reflecting the diversity of fatty acid composition of flax oil. An approach for the determination of full-length sequences of SAD1, SAD2, FAD2A, FAD2B, FAD3A, and FAD3B genes using the Illumina platform was developed and deep sequencing of the 6 genes in 84 flax samples was performed on MiSeq. The obtained high coverage (about 400x on average) enabled accurate assessment of polymorphisms in SAD1, SAD2, FAD2A, FAD2B, FAD3A, and FAD3B genes and evaluation of cultivar/line heterogeneity. The highest level of genetic diversity was observed for FAD3A and FAD3B genes – 91 and 62 polymorphisms respectively. Correlation analysis revealed associations between particular variants in SAD and FAD genes and predominantly those fatty acids whose conversion they catalyze: SAD – stearic and oleic acids, FAD2 – oleic and linoleic acids, FAD3 – linoleic and linolenic acids. All except one low-linolenic flax cultivars/lines contained both the substitution of tryptophan to stop codon in the FAD3A gene and histidine to tyrosine substitution in the FAD3B gene, while samples with only one of these polymorphisms had medium content of linolenic acid and cultivars/lines without them were high-linolenic. Conclusions: Genetic polymorphism of SAD and FAD genes was evaluated in the collection of flax cultivars and lines with diverse oil composition, and associations between particular polymorphisms and the ratio of fatty acids were revealed. The achieved results are the basis for the development of marker-assisted selection and DNA-based certification of flax cultivars. © 2020, The Author(s)