Influence of RNA interference in p53 gene on the expressions of genes involved in ultraviolet B-induced premature senescence and photocarcinogenesis in human skin fibroblasts / 中华皮肤科杂志
Chinese Journal of Dermatology
; (12): 259-262, 2014.
Article
in Zh
| WPRIM
| ID: wpr-447017
Responsible library:
WPRO
ABSTRACT
Objective To evaluate the effect of RNA interference in p53 gene on the expressions of genes involved in ultraviolet B (UVB)-induced premature senescence and photocarcinogenesis in human skin fibroblasts (HSFs).Methods A previously established HSF cell clone with repressed expression of p53,which was named as HSF-p53,was cultured and irradiated with a subcytotoxic dose (10 mJ/cm2) of UVB once a day for five consecutive days.The HSFs with normal expression of p53 served as the control.Subsequently,β-galactosidase (SA-β-gal)-staining was performed to estimate the degree of senescence,quantitative real-time PCR array was performed to determine the mRNA expressions of photocarcinogenesis-and senescence-associated genes,including p53,p21,p19,p16,pRb,fibronectin,osteonectin,smooth muscle 22 (SM22),bax,bcl-2,hypoxia-inducible factor-1 α (HIF-1α),vascular endothelial growth factor(VEGF),and human double minute-2 (hdm2).Statistical analysis was carried out by Student's t test using the software SPSS 10.0.Results The percentage of SA-β-gal-positive cells in irradiated HSF-p53 was 19.70% ± 0.85%,significantly higher than that in unirradiated HSF-p53 (12.77% ± 0.81%,t =6.45,P < 0.05),but lower than that in irradiated control HSFs (50.48% ± 5.30%,t =7.86,P < 0.05),and similar to that in unirradiated control HSFs (18.50% ± 0.45%,t =2.57,P > 0.05).Compared with the control HSFs,the HSF-p53 showed decreased expressions of p21,p19,fibronectin,osteonectin,SM22 and bax genes (all P < 0.05),but increased expressions of bcl-2,HIF-1α,VEGF and hdm2 genes (all P < 0.05),and a similar expression of p16 gene (P > 0.05); the repeated UVB radiation significantly promoted the expressions of p16 and pRb genes (both P < 0.05),but had no obvious effect on the expressions of the other genes in HSF-p53 compared with unirradiated HSF-p53 (all P > 0.05).Conclusions The inhibition of p53 expression may decelerate the UVB-induced premature senescence in HSFs,which may be involved in the p53-dependent tumor suppression.
Full text:
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Database:
WPRIM
Language:
Zh
Journal:
Chinese Journal of Dermatology
Year:
2014
Document type:
Article