Dbait molecule against acquired resistance in the treatment of cancer
A cancer treatment and molecular technology, applied in the fields of medicine and oncology, can solve the problem that the role of tumor cell subsets is not clear
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Embodiment 1
[0149] Example 1: AsiDNA sensitizes tumor cells to AsiDNA therapy
[0150] Materials and methods
[0151] cell culture
[0152] The triple-negative breast cancer cell line MDA-MB-231 was purchased from ATCC and grown according to the supplier's instructions. Briefly, MDA-MB-231 cells were grown in L15 Leibovitz medium supplemented with 10% fetal bovine serum (FBS) and maintained at 37°C in a humidified atmosphere of 0% CO2.
[0153] AsiDNA treatment and cell viability measurement
[0154] Cells were seeded at an appropriate density in 6-well culture plates and incubated at 37°C for 24h before addition of AsiDNA. Cells were harvested 7 days after treatment, stained with 0.4% trypan blue (Sigma Aldrich, Saint-Louis, USA), and counted under a microscope using Kova slides. Cell viability was calculated as the ratio of viable treated cells / viable untreated cells. Cell death was calculated as the number of dead cells among the total number of cells counted. Cells were then w...
Embodiment 2
[0162] Example 2: AsiDNA eliminates the emergence of resistance to PARP inhibitors in breast cancer
[0163] Materials and methods
[0164] cell culture
[0165] The triple-negative breast cancer cell line BC227 (BRCA2 - / - ; patient-derived cell lines from the Curie institute) were grown according to the supplier's instructions. The BC227 cell line was grown in DMEM medium supplemented with 10% FBS and 10 μg / ml insulin and maintained at 37 °C and 5% CO 2 in a humid atmosphere.
[0166] Drug treatment and cell viability measurement
[0167] For repeated cycles of the treatment regimen, drug cytotoxicity was measured by relative survival and cell death quantification. Cells were seeded in 6-well culture plates at an appropriate density and incubated at 37°C for 24 h before drug addition (olaparib 10 μM or talazoparib 0.1 μM with or without 2.5 μM AsiDNA). Cells were harvested 7 days after treatment, stained with 0.4% trypan blue (Sigma Aldrich, Saint-Louis, USA), and cou...
Embodiment 3
[0172] Example 3: AsiDNA reverses acquired resistance to talazopanib in breast cancer.
[0173] Materials and methods
[0174] cell culture
[0175] The triple-negative breast cancer cell line BC227 (BRCA2 - / - ; patient-derived cell lines from Institut Curie) were grown according to the supplier's instructions. The BC227 cell line was grown in DMEM medium supplemented with 10% FBS and 10 μg / ml insulin and maintained at 37 °C and 5% CO 2 in a humid atmosphere.
[0176] Drug treatment and cell viability measurement
[0177] For repeated cycles of the treatment regimen, talazopanib (100 nM) cytotoxicity was measured by relative viability and quantification of cell death. Cells were seeded in 6-well culture plates at an appropriate density and incubated at 37°C for 24 h before drug addition. Cells were harvested 7 days after treatment, stained with 0.4% trypan blue (Sigma Aldrich, Saint-Louis, USA), and counted under a microscope using Kova slides. Cell viability was calc...
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