Sovrem Tekhnologii Med, 2013, 5(4):19-22

The study of cisplatin effect on hydrogen peroxide and pH level in HeLa Kyoto cell line using genetically-encoded sensors

The aim of the investigation was to study the changes of hydrogen peroxide level and pH level in cytoplasm of cervical cancer cells HeLa Kyoto on cytotoxic exposure using genetically encoded sensors of hydrogen peroxide and pH. Materials and Methods. In the study we used two cell lines of human cervical cancer HeLa Kyoto containing in cytoplasm a genetically encoded sensor of hydrogen peroxide HyPer2 and a sensor pH HyPer2-C199S. To assess toxic effect of cisplatin on HeLa Kyoto cells we used a standard MTT assay. The changes of pH and hydrogen peroxide (H2O2) level were determined using fluorescence microscopy by modification in proportion between fluorescence intensities at sensor's excitation at two wavelengths: 500 and 420 nm (F500/F420). During the experiment the cells were kept in incubator at 37.0°C in carbonate-free and serum-free medium MEM. Cisplatin solution at final concentration corresponding to IC50according to MTT assay was added directly in culture medium. MEM with no cisplatin added was used as a control medium. Results. Addition of cisplatin resulted in no changes in hydrogen peroxide and pH level in cytoplasm of HeLa Kyoto cells expressing corresponding sensors during the whole period of observation (20 min). Conclusion. The use of genetically encoded sensors enables to demonstrate cisplatin to have no effect on hydrogen peroxide and pH level in HeLa Kyoto cells.

Belova AS, Mishina NM, Orlova AG, Sergeeva YA, Maslennikova AV, Brilkina AA, Shakhova NM, Belousov VV, Lukyanov SA

IBCH: 4755
Кол-во цитирований на 03.2024: 2
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