METHOD FOR DETERMINING THE REDUCTION OF RADIATION-INDUCED MIGRATION OF HUMAN BREAST CANCER CELLS OF THE MCF-7 LINE Russian patent published in 2023 - IPC A61K31/4184 A61P35/04 A61N5/10 C12M3/04 G01N33/58 C12Q1/6886 

FIELD: medicine.

SUBSTANCE: the proposed group of inventions relates to medicine, namely oncology, and can be used to determine the reduction of radiation-induced migration of human breast cancer cells of the MCF-7 line. A method is proposed in which the migration activity of tumor cells in vitro is determined using a wound healing test: cells are incubated, water-insoluble dimeric bisbenzimidazole with 5 methylene groups is added as part of a linker (DB(5) or water-insoluble dimeric bisbenzimidazole with 7 methylene groups (DB(7), after 24 hours a wound is created on a continuous cell monolayer and the remaining cells are irradiated at a dose of 4 Gy, incubated for 48 hours at a temperature of +37 ° C, the width of the cell-free band is determined 24 and 48 hours after irradiation in comparison with the initial value, accepted for 100%. The coefficient K of suppression of migration activity is calculated. With a K value equal to 26.8% after 24 hours and 19.9% after 48 hours of combined exposure to DB(5), a decrease in radiation-induced cell migration under the influence of DB(5) was recorded. At a K value of 17.9%, after 24 hours of combined exposure to DB(7), a decrease in radiation-induced cell migration under the influence of DB(7) was recorded. A method is proposed in which the migration activity of tumor cells is determined using transwell analysis: the cells are incubated at a temperature of +37 ° C with DB(5) or DB(7) for 24 hours, then the cells are irradiated at a dose of 4 Gy, after 48 hours the cells are washed from DB(5) or DB(7) and are sown in translunary inserts, incubated for three days at a temperature of +37 ° C, after which the number of migrated cells is calculated using a light microscope in comparison with single irradiation at a dose of 4 Gy. With a 2.3-fold decrease in the number of migrated cells under the influence of DB(5) and irradiation and 3.5-fold under the influence of DB(7) and irradiation, a decrease in radiation-induced cell migration was recorded compared to single irradiation at a dose of 4 Gy. A method is proposed in which the expression level of the OCLN gene is determined: cells are incubated at a temperature of +37 ° C with DB(5) or DB(7) for 24 hours, then the cells are irradiated at a dose of 4 Gy, mRNA is isolated after 48 hours, then real-time PCR is performed, threshold amplification cycles are determined (Ct), the change in the relative expression level of the OCLN gene is analyzed using the computational method of ΔΔCt. When determining an increase in the radiation-induced level of OCLN gene expression by 2.4 times under the influence of DB(5) and 1.9 times under the influence of DB(7) compared with single irradiation at a dose of 4 Gy, a decrease in radiation-induced cell migration was recorded.

EFFECT: the methods provide an effective determination of the reduction of radiation-induced migration of human breast cancer cells of the MCF-7 line by using the compound DB(5) or DB(7) in combination with ionizing radiation.

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