PMID- 20846475
OWN - NLM
STAT- MEDLINE
DCOM- 20101019
LR  - 20131121
IS  - 0393-974X (Print)
IS  - 0393-974X (Linking)
VI  - 24
IP  - 3
DP  - 2010 Jul-Sep
TI  - Specific inhibition of protein kinase Cbeta expression by antisense RNA affects 
      the activation of Jurkat T lymphoma cells.
PG  - 273-85
AB  - Antisense RNA technology was employed to specifically inhibit the expression of 
      the protein kinase Cbeta (PKCbeta) isoform in Jurkat cells, to explore its 
      influence on the expression of surface antigens (CD69) and the cytokines 
      interleukin-8 (IL-8), tumour necrosis factor (TNF)-alpha and beta, and to 
      characterise its controversial involvement in the expression of IL-2 and its 
      receptor (IL-2R). Transfection of cells with an antisense PKCbeta construct 
      (as-PKCbeta-pREP3) significantly increased IL-2R/CD25 expression in phorbol 
      12-myristate 13-acetate (PMA)-stimulated as-PKCbeta-pREP3 transfectants, in 
      contrast to Jurkat cells transfected with a control as-PKCalpha-pREP3 plasmid. 
      IL-2 production, in contrast, was strongly inhibited in both transfectant 
      populations stimulated by PMA plus the calcium ionophore ionomycin. Three clones 
      (asb1/asb2/asb3), selected from as-PKCbeta-pREP3 transfectants, showed decreased 
      PKCbeta protein levels (40 percent, 50 percent and 60 percent, respectively, as 
      determined by western blotting) and mRNA levels. The specific inhibition was 
      confirmed in immunoblots for other PKC (alpha, delta, epsilon, gamma, theta, and 
      lambda lambda/tau) isoforms and in immunoprecipitates from representative 
      (c2/asb2) clones. Stimulation of PKCbeta-depleted clones significantly increased 
      CD25 expression but decreased IL-2 production (similarly to as-PKCbeta-pREP3 
      transfectants) and IL-2 message levels. CD69 expression and IL-8 secretion were 
      significantly decreased, but TNFbeta message levels were highly increased in 
      asb2/asb3 clones, without affecting TNFalpha secretion. Analysis of the 
      mitogen-activated protein kinase (MAP Kinase) signalling pathway showed unaltered 
      extracellular signal regulated kinase 1/2 (ERK1/2) and p38 phosphorylation but 
      increased activation of c-Jun N-terminal kinase (JNK1) and its substrate, the 
      transcription factor ATF-2 (activated transcription factor-2), which are involved 
      in IL-2 gene expression. Our results revealed new PKCbeta functions, affecting 
      CD69 expression and IL-8 production, and support the requirement for PKCbeta in 
      IL-2 secretion/transcription and IL-2R regulation.
FAU - Cervino, M C
AU  - Cervino MC
AD  - Departamento de Bioquimica y Biologia Molecular, Universidad de Santiago de 
      Compostela, Santiago de Compostela, Spain.
FAU - Lopez-Lago, M A
AU  - Lopez-Lago MA
FAU - Vinuela, J E
AU  - Vinuela JE
FAU - Barja, P
AU  - Barja P
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Italy
TA  - J Biol Regul Homeost Agents
JT  - Journal of biological regulators and homeostatic agents
JID - 8809253
RN  - 0 (ATF2 protein, human)
RN  - 0 (Activating Transcription Factor 2)
RN  - 0 (Antigens, CD)
RN  - 0 (Antigens, Differentiation, T-Lymphocyte)
RN  - 0 (CD69 antigen)
RN  - 0 (IL2RA protein, human)
RN  - 0 (Interleukin-2)
RN  - 0 (Interleukin-2 Receptor alpha Subunit)
RN  - 0 (Interleukin-8)
RN  - 0 (Lectins, C-Type)
RN  - 0 (RNA, Antisense)
RN  - 0 (Receptors, Interleukin-2)
RN  - EC 2.7.11.13 (Protein Kinase C)
RN  - EC 2.7.11.13 (Protein Kinase C beta)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 8)
SB  - IM
MH  - Activating Transcription Factor 2/metabolism
MH  - Antigens, CD/analysis
MH  - Antigens, Differentiation, T-Lymphocyte/analysis
MH  - Blotting, Western
MH  - Humans
MH  - Interleukin-2/genetics
MH  - Interleukin-2 Receptor alpha Subunit/analysis
MH  - Interleukin-8/biosynthesis
MH  - Jurkat Cells
MH  - Lectins, C-Type/analysis
MH  - Lymphocyte Activation
MH  - Lymphoma, T-Cell/*immunology
MH  - MAP Kinase Signaling System
MH  - Mitogen-Activated Protein Kinase 8/metabolism
MH  - Phosphorylation
MH  - Protein Kinase C/analysis/antagonists & inhibitors/*physiology
MH  - Protein Kinase C beta
MH  - RNA, Antisense/*genetics
MH  - Receptors, Interleukin-2/genetics
MH  - Transfection
EDAT- 2010/09/18 06:00
MHDA- 2010/10/20 06:00
CRDT- 2010/09/18 06:00
PHST- 2010/09/18 06:00 [entrez]
PHST- 2010/09/18 06:00 [pubmed]
PHST- 2010/10/20 06:00 [medline]
AID - 5 [pii]
PST - ppublish
SO  - J Biol Regul Homeost Agents. 2010 Jul-Sep;24(3):273-85.