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

作　　者：Ewa AUGUSTIN Barbara BOROWA-MAZGAJ Agnieszka KIKULSKA Milena KORDALEWSKA Monika PAWLOWSKA 

机构地区：[1]Department of Pharmaceutical Technology and Biochemistry, Chemical Faculty, Gdat~sk University of Technology, Narutowicza Str1:1./12, 80-233 Gda~sk, Poland

出　　处：《Acta Pharmacologica Sinica》2013年第1期146-156,共11页中国药理学报（英文版）

摘　　要：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 （CliO） cells. Methods： Three CliO cell lines were examined： wild-type CliO 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-13-galactosidase expression analy- sis. 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 CliO, CHO-HR and CHO-HR-3A4 cells were 0.087＋0.005, 0.032＋0.0001, and 0.064＋0.0095 pmol/L, respectively. The cell cycle analysis revealed that both CliO 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 0-1305 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 CliO cells.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 （CliO） cells. Methods： Three CliO cell lines were examined： wild-type CliO 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-13-galactosidase expression analy- sis. 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 CliO, CHO-HR and CHO-HR-3A4 cells were 0.087＋0.005, 0.032＋0.0001, and 0.064＋0.0095 pmol/L, respectively. The cell cycle analysis revealed that both CliO 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 0-1305 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 CliO cells.

关 键 词：APOPTOSIS triazoloacridinones C-1305 CHO cells CYP3A4 cellular response APOPTOSIS NECROSIS SENESCENCE 

分 类 号：Q959.133[生物学—动物学] Q78