title: Expression analysis of genes involved in apoptosis, proliferation and endoplasmic reticulum stress in ionomycin/PMA treated Jurkat cells.
abstract: OBJECTIVE
  Activation of T cells by direct stimulation with phorbol 12-myristate 13-acetate (PMA) and ionomycin (Io) results in numerous downstream signals that activate pathways enabling T cells to proliferate and produce cytokines. Inducible T cell activation is regulated predominantly at the transcriptional level. Therefore, we were interested to analyze the transcriptional activity of the 19 genes involved in the regulation of several important cellular processes.
  
  
  METHODS
  Quantitative real-time (RT) PCR analysis was performed using mRNA-specific primers and SybrGreen for relative mRNA expression levels of all the examined genes.
  
  
  RESULTS
  Our results showed c-kit expression in Jurkat cells, further confirmed by sequencing of c-kit mRNAspecific PCR product. The expected increased expression of interleukin (IL)-2 mRNA, together with moderate Ki-67 upregulation, indicate the proliferation of PMA/Io treated Jurkat cells. Significant upregulation of nuclear factor (NF)-κB, JNK and the prosurvival Bcl-2 was followed by activation of only one protein kinase RNA-like endoplasmic reticulum kinase (PERK) out of 3 main endoplasmic reticulum (ER) stress subpathways (ATF6 and spliced XBP were downregulated). NF-κB and JNK activation, as well as ERK downregulation were reactive oxygen species (ROS)-independent, shown by the lack of activation of antioxidative enzymes (SOD, NOS, GSTP1, gGCS and GR). C-kit was downregulated in the absence of exogenous SCF (c-kit ligand).
  
  
  CONCLUSIONS
  Based on these data it is concluded that the PMA/Io treatment of Jurkat cells induced increased expression of IL-2, followed by upregulation of prosurvival genes belonging to the Bcl-2 family. Neither c-kit nor the antioxidative system were activated, excluding their role in Jurkat T-cell activation in the absence of exogenous c-kit ligand SCF.
authors: Clinical Center of Vojvodina, Medical Faculty, University of Novi Sad, Novi Sad, Serbia. stankov_karmen@uns.ac.rs
authors: K
authors: K
authors: Stankov
authors: G
authors: G
authors: Bogdanovic
authors: S
authors: S
authors: Stankov
authors: D
authors: D
authors: Draskovic
authors: G
authors: G
authors: Grubor-Lajsic
authors: M
authors: M
authors: Spasic
authors: D
authors: D
authors: Blagojevic
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D014408:Biomarkers, Tumor / Q000235:genetics*; D061207:Calcium Ionophores / Q000494:pharmacology; D002273:Carcinogens / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects*; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics; D015759:Ionomycin / Q000494:pharmacology*; D019169:Jurkat Cells; D016328:NF-kappa B / Q000235:genetics; D019089:Stem Cell Factor / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D015533:Transcriptional Activation / Q000187:drug effects*
chemical_list: D014408:Biomarkers, Tumor; D061207:Calcium Ionophores; D002273:Carcinogens; D007376:Interleukin-2; D016328:NF-kappa B; D019089:Stem Cell Factor; D015759:Ionomycin; D013755:Tetradecanoylphorbol Acetate