The Sensitivity of Hela Kyoto Cell Line Transfected with Sensor HyPer2 to Cisplatin

The aim of the investigation is to compare by means of MTT assay cytotoxic effect of cisplatin on the cells of HeLa Kyoto line and HeLa Kyoto line containing genetically-encoded sensor of hydrogen peroxide HyPer2 (HeLa Kyoto–HyPer2 line), and using staining by trypan blue to identify the doses of cisplatin causing cell death at different exposure time.

Materials and Methods. A HeLa Kyoto cell line of human cervical carcinoma and HeLa Kyota line transfected with the cytoplasmic sensor of hydrogen peroxide (HeLa Kyoto–HyPer2) were used in the study. The analysis of cytotoxic and antiproliferative action of cisplatin in relation to the given cells was performed using MTT assay. Cell viability was determined after 24 h of incubation with the preparation at concentrations from 0 to 50 μmol/L, then within the period from 0 to 24 h with an interval of 2 h at concentration of IC₅₀; and also after 2, 4, 6, 8 h at concentrations from 9.3 to 833.3 μmol/L a quantity of live and destructed cells was counted using staining by trypan blue.

Results. After cisplatin expose the dose-response curves for cell viability of Hela Kyoto and HeLa Kyoto–HyPer2 cell lines were built according to MTT assay data. It was established that concentration of IC₅₀ corresponding to the dose causing a loss of viability of 50% of cells is 1.3 times lower for HeLa Kyoto–HyPer2 compared to HeLa Kyoto. The results of staining by a vital agent trypan blue showed that inhibiting effects of cisplatin in concentration of IC₅₀ by 24 h are mainly linked with the delay of cell division but not with their death. At concentrations up to 52 μmol/L damage of the membranes does not occur during 8 h, and at superhigh concentrations — 416.7 μmol/L — the damage is possible already 4 h after the exposure.

Conclusion. Comparison of sensibility of the two cell lines to the effect of cisplatin showed that transfection of the cells with the fluorescent protein results in the increase of the sensitivity to cisplatin. When HeLa Kyoto–HyPer2 cells are exposed to the preparation at concentration of IC₅₀ during 24 h, inhibition of cell division is observed; higher concentrations of the preparation cause increase of the number of dead cells and diminish the terms of their destruction.

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Belova A.S., Orlova A.G., Brilkina А.А., Maslennikova A.V. The Sensitivity of Hela Kyoto Cell Line Transfected with Sensor HyPer2 to Cisplatin. Sovremennye tehnologii v medicine 2014; 6(4): 7