Medical Radiology and Radiation Safety. 2025. Vol. 70. № 6
DOI:10.33266/1024-6177-2025-70-6-12-19
A.A. Melnikova1, 2, A.A. Afonin1, L.N. Komarova1
Integrative Analysis of Pmaip1 and Birc5 Gene Expression, as Well as Protein-Protein Interactions in Sk-N-Be(2) Neuroblastoma Cells in Combination Therapy
1 National Research Nuclear University MEPhI, Obninsk, Russia
2 A.F. Tsyb Medical Radiological Research Center, Obninsk, Russia
Contact person: A.A. Melnikova, e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.
ABSTRACT
Purpose: Evaluation of the effect of ionizing radiation with various linear energy transfer on the expression of the PMAIP1 (Noxa) and BIRC5 (Survivin) genes to elucidate the molecular mechanisms determining the radiosensitivity of tumor cells.
Material and methods: The object of the study was SK-N-BE(2) cells. Four study groups were formed: a group exposed to ionizing radiation; a group treated with doxorubicin; a group of combined exposure to ionizing radiation and doxorubicin; and an untreated control group.
The cells were irradiated with 12C ions at the U-70 accelerator of the Institute for High Energy Physics (IHEP) of the National Research Centre «Kurchatov Institute» (Protvino, Moscow Region).
The irradiation was carried out at a dose of 4 Gy in an aqueous phantom with an average energy of 455 MeV/nucleon. The average LET of radiation in the initial phase was 11 keV/µm, with a peak of 120‒140 keV/µm. The cells were also irradiated with gamma radiation on the basis of the Russian Institute of Radiology and Agroecology at the unique scientific facility «Gamma radiation irradiation unit GUR-120» (radiation source 60Co, 1.25 MeV) at a dose of 4 Gy. The dose rate was 0.9 Gy /min. The cells were treated with the chemotherapy drug doxorubicin at a concentration of 0.004 mg/ml 24 hours before irradiation. Total RNA was isolated using an RNA Solo kit and quantified spectrophotometrically (NanoDrop ND-1000). Reverse transcription and amplification were performed simultaneously in real time using the OneTube RT-PCR kit with SYBR Green I as a fluorescent indicator. Key regulatory targets were identified from a set of common targets using STRING (protein-protein interaction network analysis, Homo sapiens), visualized and analyzed in Cytoscape v3.10.3 (CytoHubba plugin), and functionally annotated using Metascape (GO and KEGG pathway analysis, p<0,01).
Results: In SK-N-BE(2) cells, doxorubicin demonstrated a significant decrease in the expression of the BIRC5 gene, which is consistent with its known proapoptotic activity. The combined effect of gamma radiation (4 Gy) and doxorubicin demonstrated an additive effect (0,04). Irradiation with 12C ions demonstrated a significant decrease in BIRC5 expression with monotherapy (0,02), but a less pronounced decrease in the case of combined action with doxorubicin (0,10). Doxorubicin induced the expression of the PMAIP1 gene (0.16), this effect was synergistically enhanced when combined with gamma radiation (0.25), but was suppressed when using 12C ions (0.05), which is probably due to the activation of antiapoptotic mechanisms.
Conclusion: The results of the analysis of Gene Ontology and gene expression in SK-N-BE(2) cells confirmed the functional specialization of PMAIP1 (mitochondrial apoptosis) and BIRC5 (cell cycle regulation). Doxorubicin and gamma radiation enhanced apoptosis by affecting these genes, while 12C ions activated DNA repair, suppressing PMAIP1 and weakening the effect on BIRC5.
Keywords: 12C ion radiation therapy, gamma therapy, Bcl-2 family proteins, SK-N-BE (2), doxorubicin, BIRC5, PMAIP1, gene ontology
For citation: Melnikova AA, Afonin AA, Komarova LN. Integrative Analysis of Pmaip1 and Birc5 Gene Expression, as Well as Protein-Protein Interactions in Sk-N-Be(2) Neuroblastoma Cells in Combination Therapy. Medical Radiology and Radiation Safety. 2025;70(6):12–19. (In Russian). DOI:10.33266/1024-6177-2025-70-6-12-19
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PDF (RUS) Full-text article (in Russian)
Conflict of interest. The authors declare no conflict of interest.
Financing. The study had no sponsorship.
Contribution. A.A. Melnikova – conducting experiments, research development; A.A. Afonin – conducting experiments; L.N. Komarova – development of the research concept, scientific supervision.
Article received: 20.07.2025. Accepted for publication: 25.08.2025.