Medical Radiology and Radiation Safety. 2022. Vol. 67. № 4

Peculiarities of Human Tumor HeLa Cells Surviving and Giving
a Stable Growth After Acute X-ray Irradiation

D.V. Guryev1, 2, A.A. Tsishnatti1, 3, S.M. Rodneva1, Yu.A. Fedotov1, 2,
D.V. Molodtsova1, T.M. Blokhinа1, 2, E.I. Yashkina1, 2,
A.N. Osipov1, 2 

¹A.I. Burnasyan Federal Medical Biophysical Center, Moscow, Russia

²N.N. Semenov Federal Research Center for Chemical Physics, Moscow, Russia

³National Research Nuclear University MEPHI, Moscow, Russia

Contact person: D.V. Guryev, e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

ABSTRACT

Purpose: The evaluation of the repair efficiency of DNA double-strand breaks (DSB), proliferative activity and the yield of cytogenetic disorders in human tumor HeLa cells which survived and gave stable growth after acute irradiation at a dose of 15 Gy.

Material and methods: HeLa human tumor cell line (cervical carcinoma) was used. Cells were irradiated on an X-ray biological installation RUST-M1 (Russia), equipped with two X-ray emitters, at a dose rate of 0.85 Gy / min, a voltage of 200 kV, a total current of 10 mA, and a 1.5 mm Al filter. To obtain clones of surviving cells (HeLaRR), after acute irradiation at a dose of 15 Gy, cell cultures were incubated under standard CO₂ incubator conditions (37 °C, 5 % CO₂) for several weeks until well proliferating cells were obtained. Immunocytochemical staining of the foci of the phosphorylated H2AX protein (γH2AX) was used to quantitatively evaluate the residual foci of DNA DSB repair. The micronuclei number was assessed in cytochalasin-B cytokinesis-blocked binucleated cells stained with acridine orange with luminescence microscopy. The doubling time of the cell population was analyzed by the cell growth curves obtained by daily cell counting for five days. The cell cycle stages distribution was assessed by flow cytometry using the propidium iodide dye. All quantitative indicators of the studies were processed using the Student’s t-test for independent samples and the Kolmogorov – Smirnov test.

Results: It was revealed that acute irradiation at a high dose leads to the selection of cells with a higher reparative capacity which is confirmed by the low yield of residual foci of DNA DSB repair and MN after testing irradiation at doses of 5 and 10 Gy. A significant decrease in the proliferative activity of cells that survived after acute X-ray irradiation at a dose of 15 Gy was revealed. The doubling time of the population of unirradiated cells at the stage of exponential growth was ~18 hours while for cells that survived after irradiation at a dose of 15 Gy ~42 hours. A change in the cell cycle phases distribution was observed.

Conclusion: Thus, acute irradiation at a high dose leads to the selection of cells with a higher reparative capacity which is confirmed by the low yield of residual γH2AX foci and MN after testing irradiation at doses of 5 and 10 Gy. The decrease in proliferative activity was accompanied by a change in the cell cycle phases distribution.

Keywords: HeLa, γH2AX, micronuclei, proliferation, residual foci, DNA double-strand breaks, Х-ray

For citation: Guryev DV, Tsishnatti AA, Rodneva SM, Fedotov YuA, Molodtsova DV, Blokhinа TM, Yashkina E.I., Osipov AN. Peculiarities of Human Tumor HeLa Cells Surviving and Giving a Stable Growth After Acute X-ray Irradiation. Medical Radiology and Radiation Safety. 2022;67(4):5–9. (In Russian). DOI:10.33266/1024-6177-2022-67-4-5-9

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Conflict of interest. All authors made the same contribution to the conception, design, conduction, date analysis of the investigation and the creation of the text of the article.

Financing. The work was conducted by the research state task «Development of approaches to reduce the radioresistance of tumor stem cells» (АААА-А19-119122000097-6).

Contribution. Article was prepared with equal participation of the authors.

Article received: 17.01.2022. Accepted for publication: 15.03.2022.