[Profile of P66SHC expression and histone modifications in replicative cell senescence and oxidative-stress induced premature senescence].
OBJECTIVE
To study the profile of P66SHc expression and histone modifications in replicatively senescenct cells and oxidative-stress inducing premature senescenct cells.


METHODS
HPF cells were continuously cultured and subcultured in vitro to build replicative cellular model. HPF cells were treated with 200 pmol/L H2 O2 four times to build oxidative-stress inducing premature senescenct model. Comparative Q-PCR was utilized to investigate target gene (P66SHC, EP300, HDAC1) expressions respectively in H2O2 treated groups and normal cell groups. Then CHIP-QPCR was conducted to analyze histone modifications of P66SHC between young cells and aging cells.


RESULTS
P66SHC expression was positive correlation with H2O2 doses and population doubling level (PDL) (R = 0.909, P = 0.000; R = 0.743, P = 0.006), while EP300 was negative correlation with H2O2 (R = - 0.922, P = 0.000) and both EP300 and HDAC1 were negative with PDL (R = -0.709, P = 0.010, R = -0.599, P = 0.040). H3 histone modifications were declined in P66SHc gene regulating region. H3-Ac, H3K9-Ac and H3K4-tri-Me were dominant in the upstream region of transcriptional site (-3.0 kb) and alternative promotor (+3.8 kb).


CONCLUSIONS
P66SHC, EP300 and HDAC1 probably play a role in cellular replicative senescence and oxidative-stress inducing premature senescence. Besides, histone modification could regulate P66SHC gene expression.
Sichuan Center for Disease Control and Prevention, Chengdu 610041, China. freefresh2000@163.com
Wei
W
Xu
Zhixiong
Z
Zhuang
Jianping
J
Yang
Linqing
L
Yang
Yuling
Y
Xu
Wenjuan
W
Zhang
D019588:Aging, Premature / Q000139:chemically induced / Q000235:genetics / Q000503:physiopathology; D049109:Cell Proliferation; D016922:Cellular Senescence / Q000235:genetics*; D005347:Fibroblasts / Q000166:cytology; D020869:Gene Expression Profiling; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide; D008168:Lung / Q000166:cytology; D018384:Oxidative Stress / Q000502:physiology*; D055765:Shc Signaling Adaptor Proteins / Q000235:genetics / Q000378:metabolism*; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1
D006657:Histones; C528024:SHC1 protein, human; D055765:Shc Signaling Adaptor Proteins; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D006861:Hydrogen Peroxide