Baculovirus p35 gene greatly enhances PC12 cell's resistance against oxidative stress.
Oxidative stress is thought to be a major contributor to the progress of the Parkinson's Disease (PD) because of the high vulnerability of dopaminergic cells against oxidative stress. The present work demonstrates that with the expression of the baculovirus p35 gene, PC12 cells could gain a high resistance against oxidative toxicants, hydrogen peroxide (H(2)O(2)) and 6-hydroxydopamine (6-OHDA). The DNA fragmentation analysis showed that PC12 cells underwent apoptosis after exposure to H(2)O(2) or 6-OHDA, while PP35 cells, a p35-expressing PC12 cell line, did not. Flow cytometric analysis showed that treatment with 150 microM H(2)O(2) or 120 microM 6-OHDA for 24 h caused 52.86% or 66.36% apoptotic cell, respectively, in PC 12 cells, but only 4.26% or 5.80% in PP35 cells. The cell viability measured by 3-(4,5-dimethylthiazal-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay indicated that H(2)O(2) and 6-OHDA induced a dose-dependent cell death on PC12 cells that were greatly remitted on PP35 cells. The viability of PP35 cells was even stronger than that of PC12 cells protected by glial cell line deprived neurotrophic factor (GDNF). The surviving PP35 cells remained normal cell morphology and showed positive with tyrosine hydroxylase (TH) immunocytochemical staining. These results indicate that baculovirus p35 gene possesses remarkable ability to rescue PC12 cells from death in experimental paradigms associated with oxidative stress.
Zhuo
Ming
M
Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, 200031, Shanghai, PR China.
Yu
Fu-rong
FR
Xu
De-hua
DH
Sun
Lan-yin
LY
Liu
Xin-yuan
XY
D000818:Animals; D016923:Cell Death / Q000187:drug effects / Q000235:genetics*; D048429:Cell Size / Q000187:drug effects / Q000235:genetics; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D004298:Dopamine / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance / Q000235:genetics*; D051100:Glial Cell Line-Derived Neurotrophic Factor; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051034:Inhibitor of Apoptosis Proteins; D009414:Nerve Growth Factors / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D009498:Neurotoxins / Q000494:pharmacology; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress / Q000235:genetics*; D016627:Oxidopamine / Q000494:pharmacology; D016716:PC12 Cells; D010300:Parkinson Disease / Q000235:genetics* / Q000378:metabolism / Q000628:therapy; D051381:Rats; D014162:Transfection; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics*
C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D051034:Inhibitor of Apoptosis Proteins; D009414:Nerve Growth Factors; D009498:Neurotoxins; D016877:Oxidants; D014764:Viral Proteins; C086430:inhibitor of apoptosis, Nucleopolyhedrovirus; D016627:Oxidopamine; D006861:Hydrogen Peroxide; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine