Acta Pharmacol Sin. 2013 Jan;34(1):146-56.

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

Augustin E, Borowa-Mazgaj B, Kikulska A, Kordalewska M, Pawłowska M.

Department of Pharmaceutical Technology and Biochemistry, Chemical Faculty, Gdańsk University of Technology, Narutowicza Str. 11/12, Gdańsk, Poland. ewa.augustin@pg.gda.pl


Abstract

We examined whether the higher expression level of CYP3A4 isoenzyme influenced the cytotoxicity and the final cellular response of the antitumor triazoloacridinone derivative C-1305 in Chinese hamster ovary (CHO) cells. Three CHO cell lines were used: 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. 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 mmol/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. In summary, CYP3A4 overexpression potently enhances the cellular responses (apoptosis, necrosis and senescence) caused by C-1305 in CHO cells. In conclusions, we demonstrated for the first time that the cytotoxic activity and cellular effects induced in tumor cells by the anticancer triazoloacridinone derivative C-1305 were affected by different expression levels of metabolic enzymes. These results can be applied in the rational design of directed antitumor therapy based on differences in the expression levels of cytochrome P450 enzymes.

 

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