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

Associations between promoter polymorphisms in key genes of lipid metabolism and miogenesis and economically valuable traits in pigs

Motivation and Aim: Huge sets of gene expression pro fi les (GEP) data, in particular for farm animals, collected over the last two decades. The main aim of such data-sets is to search for gene expression differences associated with economically valuable traits. In an earlier article we showed subdivision of GEP into constitutive and variable part according to the individual changes in gene expression. Interesting, that difference in expression of genes from the second group can be possibly explained by effect of paratypic component [1]. In a present research we checked a possible in fl uence of genetic component on association between regulation of gene expression and development of economically valuable traits. It was done by association analysis of mononuclear promoter polymorphisms in key genes of lipid metabolism, control of myogenesis and meat and fat characteristics of two different industrial pig crosses. Methods and Algorithms: Polymorphic promoter regions of Acyl-CoA desaturase (SCD) [-233 T/C], Leptin (LEP) [-11606 C/T; -12109 T/A], Myoblast determination protein 1 (MYOD) [-38 G/A], Myogenic factor 6 (MYF6) [-375 T/C], Osteopontin (OPN) [-24 G/A] were genotyped with KASPar system directly from genomic DNA of two- and three-breed pig crosses of Italian selection. Phenol \u2013 chloroform extraction was used to isolate DNA from muscle tissue. Statistical analysis was carried out using the SPSS 17.0 software (SPSS statistics; 2008) Statistica 7.0 (StatSoft Inc, 2004), MedCalc Software 12.0.3, Microsoft Excel. Results: The frequencies of alleles and genotypes showed noticeable differences for all studied SNPs. Among all genes except Lep two-breed cross differ from three- breed cross with higher level of heterozygosity. Most of associations (P<0,05) were observed for promoter SNP of Opn gene and back fat, characteristics of meat production of two-breed cross. Interesting, that revealed associations of production traits and SNP genotypes in promoter region are breed and sex-dependent. Associations between all SNP genotypes and average daily gain in different periods of ontogenesis were observed only for three-breed cross pigs (P < 0,05,P < 0,01,P < 0,001). Conclusion: Received data demonstrates that associations between genotypes in promoter region of key genes of lipid metabolism, genes controlling myogenesis and production traits among analyzed groups of pigs depend on sex, genetic background (two- and three-breed crosses) and stages of development. Obviously complicated character of genetic component in fl uencing regulation of the key genes of economically valuable production traits can endow to variable part of GE