Establishment of Thiopurine S-Methyltransferase Gene Knockdown in Jurkat T-lymphocytes

• DOI: 10.1097/ftd.0b013e31826ec4b4
• Publication Date: 2012-09-12T00:37:12Z
• Authors: Misbah Misdaq; Reiner Andag; Michael Oellerich; Abdul R. Asif; Nicolas von Ahsen
• Keywords: Jurkat cells; Thiopurine methyltransferase

Background: Thiopurine S-methyltransferase (TPMT) is an excellent example of enzyme whose pharmacogenetic polymorphisms affect efficacy and toxicity a drug. The association between TPMT activity thiopurine-related myelosuppression well recognized. To study the significance deficiency in thiopurine metabolism immunosuppressive vitro, we established RNA interference-based knockdown (kd) Jurkat cell line. Results: In kd cells, expression was reduced to 73% at level 83% protein level. cells were more sensitive 6-mercaptopurine (6-MP) (10 μmol/L) 6-thioguanine (6-TG) (8 than wild-type (wt) (32% versus 20%) (18% 9%), respectively. Both wt 6-TG–induced apoptosis 6-MP. 6-TG also affected by levels 6-MP as reflected IC60, concentrations that is, [4.6 μmol/L 4.7 (kd)], [2.7 0.8 (kd)]. IC60 induced significant both (257%, 314%) with (323% 306%) 6-TG, At nucleotides (6-TGN) accumulation 518 447 pmol/million On other hand 6-TGN 477 570 6-Methylated mercaptopurine (6-MeMP) (194 10 cells) Conclusion: We conclude are appropriate vitro model investigate therapy could be helpful understanding possible clinical consequences polymorphism.