Moscow, Moscow, Russian Federation
Recently, the use of pathogenic viruses including adenoviruses to fight cancer cells, has become relevant. Cervical carcinoma cell line (HeLa) culture is a convenient model to study the factors influencing the cancer cells viability. This work aim is to study the transmembrane potentials (TMP) and mitochondria energy activity in pure HeLa cells during its defrosting and after infected with adenovirus. A polychromatic fluorescent probe 4-n-dimethylamnostyryl-1-methylpyridinium (DSM) was used to evaluate the cells cytophysical parameters. Microfluorometric studies found that high TMP ≥ | -150 | mv is reached after 6-12 hours and the maximum TMP ≥ |- 200| mv is reached after 24 hours after thawing and culturing pure cells. It was found that adenovirus reproduction in HeLa culture cells leads to mitochondria deenergization, a change in the electrical and structural properties of cell membranes and nuclear chromatin, as a result, to disruption of cancer cells viability.
HeLa cell culture, adenovirus infection, cryopreservation, transmembrane potentials, mitochondria energy
1. Podchernyaeva R.Ya., Kornilaeva G.V., Morozova G.I. Novyy podhod k metodam krio i dekonservacii dlya sohraneniya genofonda kollekcii kletochnyh kul'tur. Citologiya, 2004, t. 46, № 9, s. 837 @@Podchernyaeva R.Y., Kornilaeva G.V., Morozova G.I. New approach to cryopreservation and deconservation methods for preservation of gene pool of cell culture collection. Cytology, 2004, vol. 46, no. 9, p. 837. (In Russ.)
2. Razumov I.A., Svyatchenko V.A. Protopopova E.V., Kochneva G.V., Kiselev N.N. i dr. Ocenka onkoliticheskoy aktivnosti ryada ortopoksvirusov, adenovirusov i parvovirusov v otnoshenii kletok gliom cheloveka. Vestnik RAMN, 2013, № 12, s. 4-8. @@Razumov I.A., Svyatchenko V.A. Protopopova E.V., Kochneva G.V., Kiselev N.N. et al. Oncolytic evaluation of a number of orthopoxviruses, adenoviruses and parvoviruses against human glioma cells activity. RAMS Bulletin, 2013, no. 12, pp. 4-8. (In Russ.)
3. Toshakov V.G., Morozova G.I., Onishchenko N.A. Energy metabolism of cryopreved rad hepatocytes. Cryo-letters, 1991, no. 12, rr. 259-272.
4. Lopatina O.A., Morozova G.I., Podchernyaeva R.Ya., Firsova E.L., Danlybaeva G.A., Mihaylova G.R., Baklanova O.V. Energeticheskoe sostoyanie kletok Mpf posle kriokonservacii. Biofizika zhivoy kletki. Puschino: RAN, IBF, 2008, t. 9, s. 84-85. @@Lopatina O.A., Morozova G.I., Podchernyaeva R.Ya., Firsova E.L., Danlybaeva G.A., Mikhailova G.R., Baklanova O.V. Energy state of Mpf cells after cryopreservation. Biofizika zhivoj kletki. Pushchino: RAS, IBF, 2008, vol. 9, pp. 84-85. (In Russ.)
5. Parkhomenko T.V, Galibin O.V., Verbitskaya E.V, Tomson V.V. Evaluation of energy potential of fresh and stored bone marrow cells using a fluorescent potential-sensitive probe. Cellular Therapy and Transplantation (CTT), 2016, vol. 5, no. 2(15), pp. 60-65.
6. Morozova G.I., Dobrecov G.E., Dubure R.R., Dubur G.Ya., Golicyn V.M., Vladimirov Yu.A. Fluorescenciya zonda 4-(n-dimetilaminostiril)-1-metilpiridiniya v zhivoy kletke. Citologiya, 1981, t. 23, № 8, s. 916-923. @@Morozova G.I., Dobretsov G.E., Dubur R.R., Dubur G.Ya., Golitsyn V.M., Vladimirov Yu.A. Fluorescence of 4- (n-dimethylaminostyryl) -1-methylpyridinium probe in a living cell. Cytology. 1981, vol. 23, no. 8, pp. 916-923 (In Russ.)
7. Dobrecov G.E., Kosnikov V.V., Morozova G.I., Lihacheva L.M., Aydyraliev R.K., Vladimirov Yu.A. Izmerenie gradienta koncentracii fluorescentnogo zonda - kationa DSM na citoplazmaticheskoy i mitohondrial'noy membranah. Biologicheskie membrany, 1986, t. 3, № 3, s. 266-273. @@Dobretsov G.E., Kosnikov V.V., Morozova G.I., Likhacheva L.M., Aidaraliev R.K., Vladimirov Yu. A. Measurement of the concentration gradient of the fluorescent probe - cation DSM on the cytoplasmic and mitochondrial membranes. Biological membranes, 1986, vol. 3, no. 3, pp. 266 -273. (In Russ.)
8. Morozova G.I., Onischenko N.A., Orzhehovskaya I. G., Korobkova E. N., Polosina O. V., Bazieva F.H., Baukina O.V. Mikrofluorimetricheskiy metod identifikacii i ocenka fiziologicheskogo sostoyaniya limfocitov i neytrofilov v cel'noy nativnoy krovi s pomosch'yu fluorescentnogo zonda-kationa DSM: klinika i eksperiment. Gematologiya i transfuziologiya, 1997, t. 42, № 3, s. 43-47. @@Morozova G.I., Onishchenko N.A., Orzechowska I.G., Korobkova E.N., Polosin V.O., Baziyeva F.H., Baukina O.V. Microfluorimetric identificaition and assessment of the physiological state of lymphocytes and neutrophils in whole native blood by means of fluorescent probe-cation DSM: clinical and experimental data. Gematal. Transfuziol., 1997, vol. 42, no. 3, pp. 43-47. (In Russ.)
9. Morozova G.I., Parkhomenko T.V., Klitsenko O.A., Tomson V.V. Stimulating Effect of Erythropoietin on Thymocyte Energetics Established in vitro with Potential-sensitiv Fluorescent Probe. Biochemistry Supple-ment Series A: Membrane and Cell Biology, Moscow: 2008, vol. 1, no. 4, pp. 325-330.
10. Tarnopol'skaya O.V., Nepomnyaschaya E.M., Birbraer V.M., Tyurbeeva M.L., Makarova E.I. Sredniy membrannyy potencial kletok opuholey pyati gistotipov. Mezhdunarodnyy zhurnal prikladnyh i fundamental'nyh issledovaniy, 2014, № 10, s. 108-109. @@Tarnopolskaya O.V., Nepomnyaschaya E.M., Birbraer V.M., Tyurbeeva M.L., Makarova E.I. Average membrane potential of five histotypes tumor cells.International Journal of Applied and Basic Research, 2014, no. 10, pp. 108-109 (In Russ.)
11. Morozova G.I. Lopatina O.A., Mihaylova G.R., Danlybaeva G.A., Podchernyaeva R.Ya. Sravnitel'noe issledovanie effektov chastotno-rezonansnyh vozdeystviy i golograficheskoy informacionnoy kopii na modeli kletok mozga hor'ka s pomosch'yu potencial-chuvstvitel'nogo fluorescentnogo zonda. Biomedicinskaya radioelektronika, 2013, № 5, s. 28-34. @@Morozova G.I., Lopatina O.A., Mikhailova G.R., Danlybaeva G.A., Podchernyaeva R.Ya.Comparative study of the frequency-resonance and holographic information copy effects on ferret brain cells as model using a potential-sensitive fluorescent probe. Biomedicine Radioengineering, 2013, no. 5, pp. 28-34. (In Russ)
12. Morozova G.I., Kornilaeva G.V., Podchernyaeva R.Ya., Kulenich T.M., Bozhenko V.K. Issledovanie vliyaniya KVCh-izlucheniya millimetrovogo diapazona na membrannye struktury v kul'ture T-limfoblastoidnyh kletok s pomosch'yu fluorescentnogo zonda-kationa DSM. Biomedicinskaya radioelektronika, 2014, № 11, s. 31-38. @@Morozova G.I., Kornilaeva G.V., Podchernyaeva R.Ya., Kulenich T.M., Bozhenko V.K. Study of influence millimeter-wave EHF- irradiation on membrane structures of the T-lymphoblastoid cells culture using a fluorescent probe- cation DSM. Biomedicine Radioengineering, 2014, no. 11, pp. 31-38. (In Russ.)
13. Morozova G.I., Kozlova M.A., Akshincev A.A. Ocenka ekologicheskoy toksichnosti vodnyh rastvorov ksenobiotikov v malyh dozah na kletkah donorskoy krovi c pomosch'yu potencial-chuvstvitel'nogo fluorescentnogo zonda. Aktual'nye voprosy biologicheskoy fiziki i himii, 2019, t. 4, № 3, s. 428-424. @@Morozova G.I., Kozlova M.A., Akshintsev A.A. Evaluation of the ecological toxicity of aqueous solutions with low xenobiotics doses on donor blood cells using a potential-seneitive fluorescent probe. Aktualnye voprosy biologicheskoj fiziki i himii, 2019, vol. 4, no. 3, pp. 428-424 (In Russ)
14. Askarova K.Z., Morozova G.I., Anoshin A.A. Modeling the Accumulation Kinetics of Anionic Photo-sensitizers in Tumor Cells with Different Trans-membrane Potentials. J. Mech.Cont.& Math. Sci., Special Issue-1, March, 2019, pp. 483-490.
15. Ckulachev V.P. Energetika biologicheskih membran. M: Nauka, 1989, 123 c. @@Skulachev V. P. Energy of biological membranes. M.: Science, 1989, 123 p. (In Russ.)
16. Morozova G.I., Poletaev A.I., Borschevskaya T.A. Invertirovannyy elektrohimicheskiy potencial na yadernoy membrane kletok i ego svyaz' s kletochnoy energetikoy. Sbornik dokladov 2-go s'ezda biofizikov Rossii. M.: 1999, s. 256-257. @@Morozova G.I., Poletaev A.I., Borschevskaya T.A. Inverted electrochemical potential on the nuclear membrane of cells and its relationship with cellular energy. Reports of the 2nd Russia biophysicists Congress. M.: 1999, pp. 256-257. (In Russ.)
17. Dobrecov G.E. Fluorescentnye zondy v issledovanii kletok, membran i lipoproteinov. M.: Nauka, 1989, 277 s. @@Dobretsov G. E. Fluorescent probes in the study of cells, membranes and lipoproteins. M.: Science, 1989, 277 p. (In Russ.)
18. Beyli I.N. Statisticheskie metody v biologii. M.: Nauka, 1963, 277 s. @@Bailey I.N. Statistical methods in biology. M.: Science, 1963, 277 p. (In Russ.)
19. Kzhyshkovska Yu.G, Staheeva M.N., Litvyakov N.V. i dr. Immunnaya sistema i effektivnost' rotivoopuholevogo lecheniya. Tomsk: izdatel'stvo TGU, 2015, 130 s. @@Kzhyshkovska Yu.G., Stakheeva M.N., Litviakov N.V. et al. Immune system and the effectiveness of anticancer treatment.Tomsk: University Publishing House, 2015, 130 p. (In Russ.)
