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 МТТ 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 МЕМ. Cisplatin solution at final concentration corresponding to IC₅₀ according to МТТ 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.

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Belova А.S., Mishina N.M., Orlova А.G., Sergeeva Е.А., Maslennikova А.V., Brilkina А.А., Shakhova N.М., Belousov V.V., Lukyanov S.А. The Study of Cisplatin Effect on Hydrogen Peroxide and pH Level in HeLa Kyoto Cell Line Using Genetically-Encoded Sensors. Sovremennye tehnologii v medicine 2013; 5(4): 19