Increased transfection of the easily oxidizable GC-rich DNA fragments into the MCF7 breast cancer cell
Kostuk S. V., Mordkovich N. N., Okorokova N. A., Veiko V. P., Malinovskaya E. M., Ershova E. S., Konkova M. S., Savinova E. A., Borzikova M. A., Muzaffarova T. A., Porokhovnik L. N., Veiko N. N., Kutsev S. I.
Oxidative Medicine and Cellular Longevity
Vol.2019, Num.2348165
Опубликовано: 2019
Тип ресурса: Статья
Аннотация:
Objective. Easily oxidizable GC-rich DNA (GC-DNA) fragments accumulate in the cell-free DNA (cfDNA) of patients with various diseases. The human oxidized DNA penetrates the MCF7 breast cancer cells and significantly changes their physiology. It can be assumed that readily oxidizable GC-DNA fragments can penetrate the cancer cells and be expressed. Methods. MCF7 cells were cultured in the presence of two types of GC-DNA probes: (1) vectors pBR322 and pEGFP and (2) plasmids carrying inserted human rDNA (pBR322-rDNA and pEGFP-rDNA). pEGFP and pEGFP-rDNA contained a CMV promoter and a fluorescent protein gene EGFP. ROS generation rate, accumulation of the DNA probes in MCF7, 8-oxodG content, expression of EGFP and NOX4, and localization of EGFP, NOX4, and 8-oxodG in MCF7 were explored. The applied methods were qPCR, fluorescent microscopy (FM), immunoassay, and flow cytometry (FCA). Results. When GC-DNA is added to the cell culture medium, it interacts with the cell surface. At the site of
Ключевые слова:
Cell culture; Cell membranes; Diseases; DNA; Fluorescence; Genes; Molecular biology; Oxidation; Polymerase chain reaction; Proteins; Cell culture mediums; Cell surfaces; Expression levels; Fluorescent microscopy; Fluorescent protein; MCF-7 breast cancer cells; Plasmid transfection; Ros generations; Nucleic acids; 8 hydroxydeoxyguanosine; alkaline phosphatase; DNA fragment; endotoxin; enhanced green fluorescent protein; epidermal growth factor receptor; plasmid DNA; proteinase K; reactive oxygen metabolite; reduced nicotinamide adenine dinucleotide phosphate oxidase 4; ribosome DNA; DNA; Article; bioaccumulation; breast cancer; cell culture; cell surface; Cytomegalovirus; DNA probe; flow cytometry; fluorescence microscopy; GC rich sequence; gene insertion; genetic transfection; human; human cell; immunoassay; MCF-7 cell line; oxidation kinetics; plasmid; polymerase chain reaction; promoter region; protein expression; protein localization; breast tumor; gene vector; genetic transfection
Язык текста: Английский
ISSN: 1942-0994
Kostuk S. V. Svetlana Viktorovna 1970-
Mordkovich N. N.
Okorokova N. A.
Veiko V. P.
Malinovskaya E. M.
Ershova E. S. Elizaveta Sergeevna 1972-
Konkova M. S.
Savinova E. A.
Borzikova M. A. Mariya Aleksandrovna 1997-
Muzaffarova T. A.
Porokhovnik L. N.
Veiko N. N.
Kutsev S. I.
Костюк С. В. Светлана Викторовна 1970-
Мордкович Н. Н.
Окорокова Н. А.
Веико В. П.
Малиновскайа Е. М.
Ершова Е. С. Елизавета Сергеевна 1972-
Конкова М. С.
Савинова Е. А.
Борзикова М. А. Мария Александровна 1997-
Музаффарова Т. А.
Пороховник Л. Н.
Веико Н. Н.
Куцев С. И.
Increased transfection of the easily oxidizable GC-rich DNA fragments into the MCF7 breast cancer cell
Текст визуальный непосредственный
Oxidative Medicine and Cellular Longevity
Vol.2019 Num.2348165
2019
Статья
Cell culture Cell membranes Diseases DNA Fluorescence Genes Molecular biology Oxidation Polymerase chain reaction Proteins Cell culture mediums Cell surfaces Expression levels Fluorescent microscopy Fluorescent protein MCF-7 breast cancer cells Plasmid transfection Ros generations Nucleic acids 8 hydroxydeoxyguanosine alkaline phosphatase DNA fragment endotoxin enhanced green fluorescent protein epidermal growth factor receptor plasmid DNA proteinase K reactive oxygen metabolite reduced nicotinamide adenine dinucleotide phosphate oxidase 4 ribosome DNA DNA Article bioaccumulation breast cancer cell culture cell surface Cytomegalovirus DNA probe flow cytometry fluorescence microscopy GC rich sequence gene insertion genetic transfection human human cell immunoassay MCF-7 cell line oxidation kinetics plasmid polymerase chain reaction promoter region protein expression protein localization breast tumor gene vector genetic transfection
Objective. Easily oxidizable GC-rich DNA (GC-DNA) fragments accumulate in the cell-free DNA (cfDNA) of patients with various diseases. The human oxidized DNA penetrates the MCF7 breast cancer cells and significantly changes their physiology. It can be assumed that readily oxidizable GC-DNA fragments can penetrate the cancer cells and be expressed. Methods. MCF7 cells were cultured in the presence of two types of GC-DNA probes: (1) vectors pBR322 and pEGFP and (2) plasmids carrying inserted human rDNA (pBR322-rDNA and pEGFP-rDNA). pEGFP and pEGFP-rDNA contained a CMV promoter and a fluorescent protein gene EGFP. ROS generation rate, accumulation of the DNA probes in MCF7, 8-oxodG content, expression of EGFP and NOX4, and localization of EGFP, NOX4, and 8-oxodG in MCF7 were explored. The applied methods were qPCR, fluorescent microscopy (FM), immunoassay, and flow cytometry (FCA). Results. When GC-DNA is added to the cell culture medium, it interacts with the cell surface. At the site of