Abstract
Results of a karyological analysis of cells CHL V-79 RJK selected for their resistance to ethidium bromide (EB) causing multidrug resistance (MDR) (subline Vebr-5) were compared with data from the microfluorimetric determination of the DNA content in individual chromosomes of the karyotype. The analysis was performed at the 11th and 88th passages. Karyotyping of Vebr-5 has shown the presence of an additional genetic material (AGM) in the form of homogenously or differentially stained regions (HSR or DSR, respectively) in two chromosomes (Z1 and Z6, loci 1p29-31 and 1q26, respectively). HSR in Z6, in the site of localization of the mdr gene of the wild type had unstable length and structure, which is characteristic of the morphological markers of the amplification of genes of the mdr family. During the long cultivation of Vebr-5 in the presence of EB (88 passages), the instability of HSR (DSR) in Z6 increased. Results of a microfluorimetric analysis of Vebr-5 at the 11th passage have shown a statistically significant increase of the DNA content not only in chromosomes Z1 and Z6 marked by HSR (DSR), but also in three chromosomes (Z5, Z12, and Z13) that have no visual morphological changes. The corresponding analysis at the 88th passage has also revealed nonrandom changes in the DNA content in four more chromosomes, including an increase in Z14 and a decrease in chromosomes 8, Z7, and Z9. A decrease in the DNA content in chromosomes is considered to be the result of a partial loss of genetic material, while its increase is considered to be the result of its translocation and/or amplification. While the coefficient of the variation of the DNA content changes about 9% for large chromosomes, it amounted to about 26% for the small ones, indicating that small chromosomes to have a greater potential for instability than large chromosomes. The obtained data not only confirm, but also enlarge, the concept of the directions and character of the destabilization of the cell genetic apparatus in the process of neoplastic transformation due to MDR acquisition by these cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- AGM:
-
additional genetic material
- EB:
-
ethidium bromide
- DM:
-
double minichromosomes
- DSR:
-
differentially stained region
- HSR:
-
homogenously stained region
- MDR:
-
multiple drug resistance
References
Agafonova, N.A., Ushal, I.E., and Kudryavtsev, B.N., Method of Televisonal DNA Fluorimetry in Individual Human Chromosomes, Tsitologiya, 2002, vol. 44, no. 9, p. 858.
Bradley, G., Juranka, P.F., and Ling, V., Mechanism of Multidrug Resistance, Biochim. Biophys. Acta, 1988, vol. 948, pp. 87–128.
Bradley, G., Neik, M., and Ling, V., P-glycoprotein Expression in Multidrug-resistant Human Ovarian Carcinoma Cell Lines, Cancer Res., 1989, vol. 49, pp. 2790–2796.
Devault, A. and Gross, P., Two Members of the Mouse mdr Gene Family Confer Multidrug Resistance with Overlapping but Disting Drug Specificity, Mol. Cell Biol., 1990, vol. 10, pp. 1652–1663.
Grinchuk, T.M., Stolyar, E.A., Gornostaev, V.S., and Ignatova, T.N., Specific Changes of Chromosomes 2 and 8 and Karyotype Transformations in Murine Hepatoma Cells XXIIa Resistant to Colchicine, Struktura i funktsii kletochnogo yadra: Tez. dokl. Vsesoyuz. simp. (Structure and Functions of Cell Nucleus: Abstr. Reports All-Russia Symp.), Pushchino, 1984, p. 35.
Grinchuk, T.M., Ignatova, T.N., Efimova, E.V., and Sorokina, E.A., Karyotypical Peculiarities of the Chinese Hamster CHO-K1 Cells Resistant to Colchicine, Tsitologiya, 1986a, vol. 28, no. 1, pp. 63–68.
Grinchuk, T.M., Sorokina, E.A., and Tarunina, M.V., Peculiarities of Karyotype of Mouse Fibroblasts of the C3M10T1/2 Line Undergoing Long Selection for the Ability to Multiply in the Presence of Ethidium Bromide, Tsitologiya, 1986b, vol. 28, no. 4, pp. 422–429.
Grinchuk, T.M., Sukhikh, T.P., Sorokina, E.A., Artsibasheva, I.V., Panshina, Yu.T., Rozanov, Yu.M., Kudryavtsev, B.N., and Ignatova, T.N., Microfluorimetric Assay of DNA Content in Individual Chromosomes Confirms Cytogenetically Revealed DNA Amplifications in Structural Variants of Chromosome 1 of Chinese Hamster Cells with Stable Multiple Drug Resistance, Dokl. AN SSSR, 1986c, vol. 286, no. 3, pp. 712–717.
Grinchuk, T.M., Ignatova, T.N., Sorokina, E.A., Artsibasheva, I.V., and Panshina, Yu.T., Chromosomal Polymorphism of Mammalian Cells with Multiple Drug Resistance. I. Analysis of Karyotype of the Chinese Hamster Cells Resistant to Ethidium Bromide at Various Passages of the First Selection Step, Tsitologiya, 1988, vol. 30, no. 3, pp. 312–319.
Grinchuk, T.M., Lipskaya, L.A., Artsibasheva, I.V., Sorokina, E.A., Panshina, Yu.T., and Ignatova, T.N., Variability of Karyotype of the Chinese Hamster CHL V-79 RJK Cells Characterized by Multiple Drug Resistance Produced by Amplification of Genes of the mdr Family, Tsitologiya, 1996, vol. 38, no. 2, pp. 161–171.
Grinchuk, T.M., Pavlenko, M.A., Lipskaya, L.A., Sorokina, E.A., Tarunina, M.V., Beryozkina, E.V., Kovaleva, Z.V., and Ignatova, T.N., Resistance of Human Chronic Promyeloleukosis Cells of K562 Line to Adriamycin Correlates with Directed Genome Destabilization-Gene MDR 1 Amplification and Nonrandom Changes in Karyotype Structure, Tsitologiya, 1998, vol. 40, no. 7, pp. 652–660.
Karpishchenko, A.I., Meditsinskie laboratornye tekhnologii (Medical Laboratory Technologies), St. Petersburg: Nauka, 1999.
Kopnin, B.P., Specific Changes in Cells Resistant to Colchicine, Genetika, 1981, vol. 17, no. 2, pp. 308–317.
Kopnin, B.P., Massino, J.S., and Gudkov, A.V., Regular Pattern of Karyotypic Alterations Accompanying Gene Amplification in Djungarian Hamster Cells Study of Colchicine Adriablastin and Methotrexate Resistance, Chromosoma, 1985, vol. 95, pp. 25–36.
Ray, M. and Mohandas, T., Proposed Banding Nomenclature for Chinese Hamster Chromosomes (Cricetulus gruseus), Cytogenet., 1975, vol. 16, pp. 83U–91U.
Riordan, J.R., Deuchars, K., Kartner, N., Alon, N., Nrent, T., and Ling, V., Amplification of P-glycoprotein Genes in Multidrug-resistant Mammalian Cell Lines, Nature, 1986, vol. 316, pp. 817–819.
Roninson, I.B., Abelson, H., Housman, D.E., Hovell, N., and Varshavsky, A., Amplification of Specific DNA Sequences Correlates with Multidrug Resistance in Chinese Hamster Cells, Nature, 1984, vol. 309, pp. 626–628.
Scotto, K.W., Biedler, J.L., and Melera, P.W., Amplification and Expression of Gene Associated with Multidrug Resistance in Mammalian Cells, Science, 1986, vol. 232, pp. 751–755.
Seabright, M., A Rapid Banding Technique for Human Chromosomes, Lancet, 1971, vol. 2, pp. 971–972.
Stein, G.I., Panteleev, V.G., Povarkova, A.V., and Kudryavtsev, B.N., Possibilities of “Videotest” Image Analyzer for Performance of Microfluorimetric Studies in Cytology, Tsitologiya, 1998, vol. 40, no. 10, pp. 913–916.
Teeter, L.D., Atsumi, S.I., Sen, S., and Kuo, M.T., DNA Amplification in Multidrug, Cross-resistant Chinese Hamster Ovary Cells. Molecular Characterization and Cytogenetical Localization, J. Cell Biol., 1986, vol. 103, pp, 1159–1166.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © T.M. Grinchuk, I.E. Ushal, I.V. Artsybasheva, M.A. Pavlenko, B.N. Kudryavtsev, 2007, published in Tsitologiya, Vol. 49, No. 6, 2007.
Rights and permissions
About this article
Cite this article
Grinchuk, T.M., Ushal, I.E., Artsybasheva, I.V. et al. Microfluorimetric analysis of dynamics of genomic changes in Chinese hamster fibroblasts CHL V-79 RJK with multiple drug resistance. Cell Tiss. Biol. 1, 518–523 (2007). https://doi.org/10.1134/S1990519X07060089
Received:
Issue Date:
DOI: https://doi.org/10.1134/S1990519X07060089