Original Article

CYP3A4 overexpression enhances the cytotoxicity of the antitumor triazoloacridinone derivative C-1305 in CHO cells

Authors: Ewa Augustin, Barbara Borowa-Mazgaj, Agnieszka Kikulska, Milena Kordalewska, Monika Pawłowska
DOI: 10.1038/aps.2012.132

Abstract

Ewa AUGUSTIN*, Barbara BOROWA-MAZGAJ, Agnieszka KIKULSKA, Milena KORDALEWSKA, Monika PAWŁOWSKA

Department of Pharmaceutical Technology and Biochemistry, Chemical Faculty, Gdańsk University of Technology, Narutowicza Str 11/12, 80–233 Gdańsk, Poland

Aim: To examine how the higher expression level of CYP3A4 isoenzyme influenced the cytotoxicity of the antitumor triazoloacridinone derivative C-1305 in Chinese hamster ovary (CHO) cells.

Methods: Three CHO cell lines were examined: wild-type CHO cells; CHO-HR cells with overexpression of human cytochrome P450 reductase (CPR); and CHO-HR-3A4 cells with coexpression of human CYP3A4 and CPR. Cellular responses caused by C-1305 were monitored using DAPI staining, cell cycle analysis, phosphatydilserine externalization analysis and SA-β-galactosidase expression analysis. Cell viability was assessed with simultaneous FDA and PI staining.

Results: Treatment with C-1305 for 72 h exhibited different levels of cytotoxicity in the 3 cell lines, and the values of IC80 in CHO, CHO-HR and CHO-HR-3A4 cells were 0.087±0.005, 0.032±0.0001, and 0.064±0.0095 μmol/L, respectively. The cell cycle analysis revealed that both CHO and CHO-HR cells underwent transient G2/M arrest, whereas CHO-HR-3A4 cells did not accumulate in this phase. Prolonged exposure up to 120 h caused time-dependent increase in the sub-G1 fraction in all the 3 cell lines. Treatment with C-1305 caused cell death through apoptosis and necrosis. However, these processes were more pronounced in the transfected CHO cells than in the wild-type cells. The cells surviving after C-1305 exposure underwent senescence.

Conclusion: CYP3A4 overexpression potently enhances the cellular responses (apoptosis, necrosis and senescence) caused by C-1305 in CHO cells.


Keywords: apoptosis; triazoloacridinones; C-1305; CHO cells; CYP3A4; cellular response; apoptosis; necrosis; senescence

This work was supported by R&D grant No 014668/009 from the Chemical Faculty of the Gdańsk University of Technology. We thank Dr Thomas FRIEDBERG and Dr Roland C WOLF from the Biomedical Research Centre, Dundee, Scotland, for providing the CHO cell lines for this study. We also thank Prof Zofia MAZERSKA and Prof Jerzy KONOPA from our department for helpful suggestion.
* To whom correspondence should be addressed.
E-mail ewa.augustin@pg.gda.pl
Received 2012-06-08 Accepted 2012-08-17
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