Tant and that none showed mutations in the kinase domain of BCR-ABL1. We identified the PI3K E545G mutation in cell line KCl-22 [11] as well as the mutationally inactivated PI3K-inhibitor PTEN (Phosphatase and tensin homolog) in the MHH-TALL-1 cell line (Figure S2) as potential causes for constitutive activity on the PI3K/AKT pathway conferring TKI resistance to these cells. Within this study, we found that overexpression with the anti-apoptotic protein MDM2 contributes to TKI resistance and that suppression of MDM2 restores sensitivity to TKI. In conclusion, we employed Ph+ cell lines JURL-MK2 and SUPB15 to investigate mechanisms of TKI resistance and sensitization. TKI nilotinib effectively induced cell apoptosis in JURL-MK2, but not in SUP-B15 cells. Inhibition of BCR-ABL1 by nilotinib and simultaneous down-regulation of your antiapoptotic protein MDM2 by BEZ235 synergized in inducing apoptosis in SUP-B15 cells (Figure six). BEZ235 operated via blocking the translational machinery as evidenced by dephosphorylation of S6 and 4E-BP1. Taken collectively, these outcomes suggest that MDM2 might be a therapeutic target to enhance TKI-mediated apoptosis and that combining PI3K/mTOR inhibitor and TKI might prove an effective novel therapeutic technique in TKI-resistant BCR-ABL1 positive leukemia.Supporting InformationFigure S1. BEZ235 inhibits phosphorylation of BCR-ABL1 downstream signaling molecule ERK but not STAT5 in SUP-B15 cells. (A) SUP-B15 cells had been respectively treated with 100 nM Imatinib, 100 nM nilotinib, 100 nM Everolimus or two M BEZ235 for 24 h. p-ERK and ERK protein levels had been analyzed by Western blot. (B) P-STAT5 and STAT5 protein expression in SUP-B15 cells were tested by Western blot after 24 h therapy with different concentrations of nilotinib and BEZ235. (TIF) Figure S2. Genomic silencing of PTEN in MHH-TALL-1. (A) Fluorescence in situ hybridization evaluation showed intact IKZF1 (left) but loss of your genomic region hosting PTEN (right) covering much of BAC clone RPM1-383D9 (red) and all of the neighboring clone RPMI-879E1. Standard cytogenetic evaluation showed no proof of genomic loss, therefore indicating focal microdeletion of the centromeric part of chromosome band 10q23.31 (not shown). (B) Genomic PCR confirmed the hemizygous loss of PTEN in cell line MHH-TALL1. (C) The remaining PTEN allele had a a single base pair insertion leading to a premature cease just after amino acid 241, evidenced by sequencing from the RT-PCR solution of cell line MHH-TALL1. (D) Cell line MHH-TALL1 did not express the PTEN protein according to Western blot evaluation. T, T-cell; B, B-cell; M, myeloid; r, resistant; s, sensitive; n.4,6-Dibromopicolinic acid site d.6-Bromo-4-chloropyridin-2-amine Formula , not accomplished.PMID:25016614 (TIF)AcknowledgementsWe are grateful to Novartis Pharmaceuticals (Basel, Switzerland) which provided nilotinib and BEZ235 for the study.Author ContributionsConceived and developed the experiments: JD HQ. Performed the experiments: JD JR MZ SN RM. Analyzed the data: JD HQ RM. Contributed reagents/materials/analysis tools: SN HD. Wrote the manuscript: JD HQ.
Redox Biology 1 (2013) 448?Contents lists offered at ScienceDirectRedox Biologyjournal homepage: elsevier/locate/redoxMini ReviewRenoprotective effect on the antioxidant curcumin: Recent findingsJoyce Trujillo a, Yolanda Irasema Chirino b, Eduardo Molina-Jij c, Ana Cristina And ica-Romero a, Edilia Tapia d, Jos?Pedraza-Chaverr?a,nDepartment of Biology, Facultad de Qu ica, UNAM, Ciudad Universitaria, 04510 M ico, DF, Mexico Unidad de Biomedicina, Facultad de Estudios Superiores Iztacala,.
