Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Aug 4:12:912741.
doi: 10.3389/fonc.2022.912741. eCollection 2022.

Reparative properties of human glioblastoma cells after single exposure to a wide range of X-ray doses

Galina Pavlova  1   2   3 , Alexandra Belyashova  1 , Ekaterina Savchenko  1 , Dmitri Panteleev  2 , Dzhirgala Shamadykova  2 , Anna Nikolaeva  1 , Svetlana Pavlova  2 , Alexander Revishchin  2 , Denis Golbin  1 , Alexander Potapov  1 , Natalia Antipina  1 , Andrey Golanov  1
Affiliations

Reparative properties of human glioblastoma cells after single exposure to a wide range of X-ray doses

Galina Pavlova et al. Front Oncol. .

Abstract

Radiation therapy induces double-stranded DNA breaks in tumor cells, which leads to their death. A fraction of glioblastoma cells repair such breaks and reinitiate tumor growth. It was necessary to identify the relationship between high radiation doses and the proliferative activity of glioblastoma cells, and to evaluate the contribution of DNA repair pathways, homologous recombination (HR), and nonhomologous end joining (NHEJ) to tumor-cell recovery. We demonstrated that the GO1 culture derived from glioblastoma cells from Patient G, who had previously been irradiated, proved to be less sensitive to radiation than the Sus\fP2 glioblastoma culture was from Patient S, who had not been exposed to radiation before. GO1 cell proliferation decreased with radiation dose, and MTT decreased to 35% after a single exposure to 125 Gγ. The proliferative potential of glioblastoma culture Sus\fP2 decreased to 35% after exposure to 5 Gγ. At low radiation doses, cell proliferation and the expression of RAD51 were decreased; at high doses, cell proliferation was correlated with Ku70 protein expression. Therefore, HR and NHEJ are involved in DNA break repair after exposure to different radiation doses. Low doses induce HR, while higher doses induce the faster but less accurate NHEJ pathway of double-stranded DNA break repair.

Keywords: Ku70; Ku80; Rad51; human glioblastoma cell cultures; radiation therapy.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Characteristics of glioblastoma cell cultures.
Figure 2
Figure 2
Brain MRI, T2 FLAIR, and T1 images of patients S (A) and G (B). There are the same typical heterogenic changes of the signal with a ring-like contrast enhancement and a signal typical for edema around the tumors.
Figure 3
Figure 3
Neurospheres of glioblastoma cell culture of GO1. (A) antibody staining for Vimentin. (B) staining for bisbenzimid, (C) antibody staining for Nestin, (D) neurospheres in phase contrast. Scale bar is 20 μm.
Figure 4
Figure 4
Neurospheres of glioblastoma cell culture of Sus\fP2. (A) antibody staining for Vimentin. (B) staining for bisbenzimid, (C) antibody staining for Nestin, (D) neurospheres in phase contrast. Scale bar is 20 μm.
Figure 5
Figure 5
Relationship between the number of proliferating cells and radiation dose according to the MTT assay. (A) Sus\fP2 cells; (B) GO1 cells, (C) Sus\fP2 cells exposed to radiation below 5 Gγ. Each value is the mean of ≥3 independent experiments ± Standard Error of the Mean (SEM). Control values on (B, C) (denoted with *) significantly exceeded the experimental ones at any radiation doses with p<0.01 according to the Mann-Whitney test.
Figure 6
Figure 6
Analysis of apoptosis and necrosis in Sus\fP2 and GO1 cell cultures exposed to 200 Gγ.
Figure 7
Figure 7
The dependence of RAD51 mRNA and protein levels on radiation dose in GO1 (A–C) and Sus\fP2 (D–F) glioblastoma cell cultures. (A, D) mRNA expression; (B, E) Western blotting; (C, F) Quantification RAD51 protein levels by ImageJ.
Figure 8
Figure 8
Expression of XRCC6 gene (A) and its product Ku70 (B, C) as a function of radiation dose in glioblastoma GO1 cell culture. Expression of XRCC5 gene (D) and its product Ku80 (E, F) as a function of radiation dose in GO1 cell culture. Expression of XRCC6 gene (G) and its product Ku70 (H, I) as a function of a single radiation dose from 5 to 250 Gγ in glioblastoma Sus\fP2 cell culture. Expression of XRCC5 gene (J) and its product Ku80 (K, L) as a function of a single radiation dose from 5 to 250 Gγ in Sus/fP2 cells.
Figure 9
Figure 9
Dose-effect relationship between Sus\fP2 (A) and GO1 (B) proliferative activity and expression of RAD51, Ku70, and Ku80 proteins.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

  • Glioblastoma Therapy: Past, Present and Future.
    Obrador E, Moreno-Murciano P, Oriol-Caballo M, López-Blanch R, Pineda B, Gutiérrez-Arroyo JL, Loras A, Gonzalez-Bonet LG, Martinez-Cadenas C, Estrela JM, Marqués-Torrejón MÁ. Obrador E, et al. Int J Mol Sci. 2024 Feb 21;25(5):2529. doi: 10.3390/ijms25052529. Int J Mol Sci. 2024. PMID: 38473776 Free PMC article. Review.
  • GQIcombi application to subdue glioma via differentiation therapy.
    Kolesnikova V, Revishchin A, Fab L, Alekseeva A, Ryabova A, Pronin I, Usachev DY, Kopylov A, Pavlova G. Kolesnikova V, et al. Front Oncol. 2024 Jun 26;14:1322795. doi: 10.3389/fonc.2024.1322795. eCollection 2024. Front Oncol. 2024. PMID: 38988707 Free PMC article.

References

    1. Ostrom QT, Bauchet L, Davis FG, Deltour I, Fisher JL, Langer CE, et al. . The epidemiology of glioma in adults: a "state of the science" review. Neuro-Oncology (2014) 16:896–913. doi: 10.1093/neuonc/nou087 - DOI - PMC - PubMed
    1. Balbous A, Cortes U, Guilloteau K, Rivet P, Pinel B, Duchesne M, et al. . A radiosensitizing effect of RAD51 inhibition in glioblastoma stem-like cells. BMC Cancer (2016) 16:604. doi: 10.1186/s12885-016-2647-9 - DOI - PMC - PubMed
    1. Helleday T, Petermann E, Lundin C, Hodgson B, Sharma RA. DNA Repair pathways as targets for cancer therapy. Nat Rev Cancer (2008) 8:193–204. doi: 10.1038/nrc2342 - DOI - PubMed
    1. Lim YC, Roberts TL, Day BW, Stringer BW, Kozlov S, Fazry S, et al. . Increased sensitivity to ionizing radiation by targeting the homologous recombination pathway in glioma initiating cells. Mol Oncol (2014) 8:1603–15. doi: 10.1016/j.molonc.2014.06.012 - DOI - PMC - PubMed
    1. Short SC, Giampieri S, Worku M, Alcaide-German M, Sioftanos G, Bourne S, et al. . Rad51 inhibition is an effective means of targeting DNA repair in glioma models and CD133+ tumor-derived cells. Neuro-Oncology (2011) 13:487–99. doi: 10.1093/neuonc/nor010 - DOI - PMC - PubMed