[Inhibition effects of par-4 antisense oligodeoxynucleotide on apoptosis of PC12 cell induced by glutamate is mediated by ERK1/2].
OBJECTIVE
To investigate the inhibition effects of par-4 antisense oligodeoxynucleotide on apoptosis of PC12 cell induced by glutamate and its signal transduction mechanism.


METHODS
(1) Cationic lipid-mediated par-4 antisense oligodeoxynucleotide (Par-4-AS-ODN) was transfected into PC12 cells before they were treated with glutamate. Mismatch oligodeoxynucleotide (MS-ODN) were also transfected into cells as controls. (2) Morphological observation and the detection of anti-apoptosis effects of par-4-AS-ODN on PC12 cells were done with the Laser Scanning confocal Microscope by double staining the cells with acridine orange/ethidium bromide (AO/EB), addition to with flow cytometry. (3) Western blot was used to detect the protein levels of par-4 and phosphorylated ERK(1/2) at threonine-202 and Tyrosine-204.


RESULTS
(1) Par-4-AS-ODN significantly suppressed up-regulation of the par-4 protein levels induced by glutamate in PC12 cells. (2) Par-4-AS-ODN could resist the decrease of phosphorylated ERK(1/2) (Thr202/Tyr204) induced by glutamate in PC12 cells. (3) Par-4 AS-ODN could inhibit apoptosis of PC12 cells induced by glutamate. But its inhibition effect could be eliminated by PD98059, a selective MEK(1) inhibitor which could inhibit phosphorylation of ERK(1/2).


CONCLUSIONS
Par-4 AS-ODN may inhibit apoptosis of PC12 cells induced by glutamate, and its inhibition effects may be medicated by the activation of ERK(1/2).
Lu
Chao
C
Department of Pediatrics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
Chen
Ji-qing
JQ
Wu
Sheng-hua
SH
Zhao
Fei
F
Chi
Xia
X
Pan
Xiao-qin
XQ
Fei
Li
L
Guo
Mei
M
Huang
Song-ming
SM
Guo
Xi-rong
XR
Chen
Rong-hua
RH
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000494:pharmacology*; D015536:Down-Regulation; D018698:Glutamic Acid / Q000494:pharmacology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000096:biosynthesis / Q000235:genetics; D048052:Mitogen-Activated Protein Kinase 3 / Q000096:biosynthesis* / Q000235:genetics; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D016716:PC12 Cells; D051381:Rats; D015398:Signal Transduction
D051017:Apoptosis Regulatory Proteins; D020319:Oligodeoxyribonucleotides, Antisense; C101296:prostate apoptosis response-4 protein; D018698:Glutamic Acid; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3