Biopolym. Cell. 1999; 15(1):28-37.
http://dx.doi.org/10.7124/bc.000502
Structure and Function of Biopolymers
The recombinant DNA pGins that contains expressing human preproinsulin gene has no mutagenic activity
1Lukash L. L., 1Podolskaya S. V., 1Evseeenko A. A., 1Suhorada He. M., 1Neborachko L. N., 1Varsanova I. S., 1Kochubei T. P.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

Uniike recombinant plasmids pBR322ins u pAins the plasmid DNA pGins at the concentration of 10 ng/ml does not manifest mutagenic activity in. somatic mammalian cells. The at>sence of pGins mutagnnic activity is confirmed in different test-systems in vitro and in vivo and presumably is determined by the increasing of the expression and the functions of the preproinsulin gene under the action, of virus regulation elements. It is shown that the preliminary treatment of somatic mammalian cells by insulin preparation protects them against the mutagenic action of recombinant plasmid pHB320, containing the genome of virus hepatitis B. The data obtained here testify to the participation of the expressing gene coding for insulin in the maintenance of genetic stability or cellular populations variation.
Full text: (PDF, in Russian)

References

[1] Kordyum VA, Frolkis VV, Lukash LL, Shulzhenko VN, Shpilevaya SP, Kostetsky IE, Titok TG, Varsanova IS, Likhacheva LI, Irodov DM. Gene therapy of mass pathologies. Biopolym. Cell. 1993;9(4):63-104
[2] Lukash LL, Neborachko LN, Sukhorada EM, Usenko EV, Varsanova IS, Podolskaya SV, Buzhievskaya TI, Kordium VA. Human insulin gene expression in cultured mammalian cells. Biopolym. Cell. 1990;6(1):82-87
[3] Explore the possibility of transforming and mutagenic effects of genetically engineered constructs containing valuable real human genes: Research report (final). Institute of Molecular Biology and Genetics. GR 01.88.0081890: Inv. N. Kiev, 1992. 97 p.
[4] Rubashevskii EL, Patskovskii IuV, Lysenko EF, Kirilenko SD, Lukash LL. The mutagenic effect of the recombinant plasmid pBR322ins carrying a native and a modified human insulin gene. Tsitol Genet. 1993;27(4):44-51.
[5] Lukash LL, Podolskaya SV, Suhorada HM, Kostetskaya KV, Kostetsky IE, Varzanova IS, Patskovsky YuV, Vavilina IV, Deis SV. The influence of alkylating agent MNNG on mutagenic effect of exogeneous recombinant DNA. Biopolym. Cell. 1995;11(1):87-91
[6] Rubashevsky EL, Patskovsky YuV, Kirilenko SD, Varzanova IS, Bogolyubova OM, Lysenko OF, Lukash LL. DNA recombinant molecules carrying different allels of human preproinsulin gene as mutagens in Chinese hamster cells. Biopolym. Cell. 1996; 12(5):83-92
[7] Kozlovskaia TM, Pumpen PP. Structure and expression of human hepatitis B virus surface antigen. Mol Biol (Mosk). 1986;20(4):884-901.
[8] Titok TG, Kostetsky IE, Tchaikovskaya TL, Varzanova IS, Kochubey TP, Lukash LL, Kordium VA. Transplantation of human preproinsulin gene into the insuline dependent diabetic and normal rats. Biopolym. Cell. 1993;9(1):39-44
[9] Lukash LL, Podolskaya SV, Neborachko LN. et al. Expression of insulin gene introduced into embryonic human cells. Proc. of the 1994 Miami Bio. Technol. Eur. Symp Short reports. Monaco, 1994:62.
[10] Maniatis T., Fritsch E. F., Sambrook J. Molecular cloning: a laboratory manual New York: Cold Spring Harbor Lab. publ., 1982 545 p.
[11] van der Eb AJ, Graham FL. Assay of transforming activity of tumor virus DNA. Methods Enzymol. 1980;65(1):826-39.
[12] Buzhievskaya TI, Lukash LL, Podolsky Eksperimental model for the study of mutagenesis and transformation induced by viruses and nucleic acids. Molecular biology techniques. Naukova Dumka, 1986:147-158.
[13] Moorhead PS, Nowell PC, Mellman WJ, Battips DM, Hungerford DA. Chromosome preparations of leukocytes cultured from human peripheral blood. Exp Cell Res. 1960;20:613-6.
[14] G. McGregor, J. Varley Methods of working with animal chromosomes. New York: Wiley, 1986. 272.
[15] Monakhov AS. Micromethod of culturing the lymphocytes from rabbit and white rat peripheral blood for chromosome study. Tsitologiia. 1982;24(2):233-8.
[16] Neborachko L. M., Varzanova I. S., Podol's'ka S. V. Gene therapy of insulin-dependent diabetes mellitus. 11 Materials Congress of Medical Geneticists Ukraine (Lviv 18-20 October 1995). Lviv, 1995:104-105.
[17] Plokhinskiy N. A. Algorithms biometrics. M.: Moscow State University Press, 1980. 150p.
[18] Shapiro NI, Varshaver NB. Rate of the spontaneous mutation process in mammalian somatic cells and several related questions. Genetika. 1976;12(7):132-49.
[19] Lukash LL, Buzhievskaya TI. Role of early visuses genes in mutagenesis. Biotechnol. Curr. Progr. Lancaster: Techn. Publ. Co., 1991:119—131.
[20] Buzhievskaya TI, Lukash LL, Rubashevsky EL. Mutagenic and transforming action of some viruses, genes and recombinant molecules. Biotechnology (PA 17604, USA). Lancaster : Curr. progr. techn. publ. Co., 1991: 133-156.
[21] Schramayr S, Caporossi D, Mak I, Jelinek T, Bacchetti S. Chromosomal damage induced by human adenovirus type 12 requires expression of the E1B 55-kilodalton viral protein. J Virol. 1990;64(5):2090-5.
[22] Baron EM, Bobrysheva IV, Varshaver NB.The plasmid pEJ6.6 carrying the activated c-Ha-ras-1 oncogene increases the mutation frequency in Chinese hamster cells. Genetika. 1992;28(6):35-40.
[23] Bobrysheva I. V. Activated cellular oncogenes c-Ha-rasl and c-myc as mutagens: genetic and molecular biological research Avtoref. dis. ... kand. biol. nauk. M.: Med. genet. tsentr RAMN, 1995. 20 s.
[24] Luaky M., Botchan . Inhibition of SV40 replication in simian cells by specific pBR322 DNA sequences. Nature. 1981. 293:79—81.
[25] Peden KW, Pipas JM, Pearson-White S, Nathans D. Isolation of mutants of an animal virus in bacteria. Science. 1980;209(4463):1392-6.
[26] Cordeli B, Goodman IM. Evolution of the mammalian insulin gene. Genes and tumor genes. New York: Raven press, 1982:115—119.
[27] Mitanchez D1, Doiron B, Chen R, Kahn A. Glucose-stimulated genes and prospects of gene therapy for type I diabetes. Endocr Rev. 1997;18(4):520-40.
[28] Hayashi M, Taguchi M, Fujita T. The IRI release from insulin gene-transduced hepatic cells responds to ambient glucose concentration. Biochem Biophys Res Commun. 1997;233(2):470-5.
[29] Makino H, Manganiello VC, Kono T. Roles of ATP in insulin actions. Annu Rev Physiol. 1994;56:273-95.
[30] Laub O, Rutter WJ. Expression of the human insulin gene and cDNA in a heterologous mammalian system. J Biol Chem. 1983;258(10):6043-50.
[31] Docherty K. Gene therapy for diabetes mellitus. Clin Sci (Lond). 1997;92(4):321-30.
[32] Lukash LL, Karpova IS, Miroshnichenko OS, Tikhonova TN, Lylo VV, Man'ko VG, Sukhorada EM, Golynskaia EL. The effect of the lectin from Sambucus nigra inflorescences on spontaneous and alkylating agent-induced mutagenesis in mammalian somatic cells. Tsitol Genet. 1997;31(5):52-60.
[33] Ranchialis VP, Bal'chiunene LS. Paradoxical effects of thiol compounds. Vestn Ross Akad Med Nauk. 1995;(1):44-9.
[34] Delrez M, Firket U. Action paradoxale d'un radioprotecteur sur la mitose et les chromosomes, in vitro. Biochem Pharmacol. 1968;17(9):1893-9.
[35] Rosin MP, Stich HF. The inhibitory effect of cysteine on the mutagenic activities of several carcinogens. Mutat Res. 1978;54(1):73-81.
[36] Stark AA, Zeiger E, Pagano DA. Glutathione mutagenesis in Salmonella typhimurium is a gamma-glutamyltranspeptidase-enhanced process involving active oxygen species. Carcinogenesis. 1988;9(5):771-7.
[37] Holme JA, Søderlund EJ, Brunborg G, Omichinski JG, Bekkedal K, Trygg B, Nelson SD, Dybing E. Different mechanisms are involved in DNA damage, bacterial mutagenicity and cytotoxicity induced by 1,2-dibromo-3-chloropropane in suspensions of rat liver cells. Carcinogenesis. 1989;10(1):49-54.
[38] Seredenin SV, Durnev AD pharmacological protection of the genome. M.: VINITI, 1992, 160p.
[39] Zasukhina GD, Sinel'shchikova TA. Mutagenesis, antimutagenesis, DNA repair. Vestn Ross Akad Med Nauk. 1993;(1):9-14
[40] Timchenko OI, Antipenko EN. Conditions for using thyroxine as an antimutagen after whole-body x-ray irradiation. Radiobiologiia. 1981;21(2):204-10.
[41] Rushkovskii SR, Chegrinets SE, Bezrukov VF, Khrapunov SN. The effect of thymogen on the chromosome aberration level in a culture of human peripheral blood lymphocytes. Tsitol Genet. 1996;30(5):81-5.
[42] Semenov VV. Mutagenesis, antimutagenesis and the cellular regulatory system. Vestn Ross Akad Med Nauk. 1995;(1):41-4.
[43] Tomilin NV genetic stability of the cells. Leningrad: Nauka, 1983. 158p.
[44] Popov LS, Gorbunova LV, Varshaver NB, Shapiro NI. Integration of SV40 DNA into the cell genome and viral mutagenesis. Genetika. 1986;22(9):2213-9.
[45] Gorbunova LV, Varshaver NB, Marshak MI, Shapiro NI. The role of the transforming A gene of SV40 in the mutagenic activity of the virus. Mol Gen Genet. 1982;187(3):473-6.
[46] Drize OB, Sokova OI, Nikashina EB, Shliankevich MA, Shapot VS. Possible role of the T antigen in inducing chromosome aberrations in SV40 virus-transformed cells. Tsitologiia. 1985;27(1):76-82.
[47] Bobrysheva IV, Varshaver NB. Characteristics of mutants induced by the c-Ha-ras1 oncogene and the nature of the oncogene's mutagenic action. Genetika. 1995;31(12):1598-604.