Transfection of HL-60 cells with CYP3A5 gene induces drug-resistant phenotype
Wang, T.; Chen, F-yuan.; Gu, C-hong.; Zhong, H.; Teng, Y.; Ouyang, R-rong.
Zhonghua Zhong Liu Za Zhi 27(8): 461-464
2005
ISSN/ISBN: 0253-3766 PMID: 16188140 Document Number: 585683
Objective To investigate if CYP3A5 gene is involved ill the molecular mechanisms for multiple drug resistance ill leukemia cells. Methods A full length cDNA of CYP3A5 gene was cloned, and a recombinant eukaryotic expression plasmid was constructed, then stably transfected cell lines were established. Furthermore, the sensitivity of those cell lines to several anticancer drugs were assessed by WIT and FCM assay. Results The recombinant plasmid was; designated as pcDNA3-CYP3A5. Transfecting HL-60 cells (which didn't show transcript of CYP3A5 gene) with recombinant plasmid pcDNA3-CYP3A5 generated HL-60/CYP3A5 cell line, and transfecting of HL-60 cells with the parental pcDNA3 vector served as control HL-60/pc; cell line. Daunorubicin induced remarkable apoptosis peaks in HL-60 and HL-60/pc cells, while such effect did not occur in HL-60/CYP3A5 cells (apoptosis cell percentage were 7.3%, 6.3% and 1.2%, respectively). Compared villi HL-60 mid HL-60/pc cells, HL-60/CYF3A5 cells were statistically significantly resistant to daunorubicin, aclacinomycin A, vincristine and harringtonine (resistance multiples were 2.89, 2.01, 4.05 and 2.79 times, respectively, P < 0.05), however the sensitivity to teniposide didn't change (resistance multiple was 1.04 times). Conclusion Transcription of CYP3A5 gene, ill leukemia cells directly induces resistance to anthracyclines and alkaloids, however die cells are still sensitive to epipodophyllotoxins. Therefore, our findings confirmed a new mechanism of multidrug resistance.