ABSTRACT
Objective To measure the expressions of miR-203 and its downstream target genes p63 and survivin in psoriasis vulgaris lesions.Methods Tissue specimens were collected from lesions of 30 patients with psoriasis vulgaris and normal skin of 30 healthy human controls.Real-time PCR was performed to detect miR-203 mRNA expression with U6 as the internal control,as well as p63 and survivin mRNA expressions with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the internal control,and Western blot to measure the protein expressions of miR-203 targets p63 and survivin.Statistical analysis was carried out by t test for intergroup comparisons and by Pearson correlation analysis for the analysis of correlation between miR-203,p63 and survivin expressions in psoriasis vulgaris lesions.Results Compared with the normal control skin,psoriasis vulgaris lesions showed significantly decreased mRNA expression level (2-△△C△△Ct) of miR-203 (0.41 ± 0.11,t =3.16,P ＜ 0.05),but increased mRNA expression levels of p63 (4.79 ± 0.63,t =4.72,P＜ 0.05) and survivin (3.43 ± 0.46,t =4.35,P＜ 0.05).The protein expression levels of p63 and survivin in these lesions were (2.40 ± 0.23) times (t =3.87,P ＜ 0.05) and (3.49 ± 0.14) times (t =4.36,P ＜ 0.05) those in the normal control skin respectively.In psoriasis vulgaris lesions,the mRNA expression level of miR-203 showed a significantly negative correlation with that of survivin (r =-0.36,P ＜ 0.05) and p63 (r =-0.43,P ＜ 0.05).Conclusion miR-203 and its downstream target genes p63 and survivin may participate in the occurrence of psoriasis vulgaris.
ABSTRACT
Objective To clarify the relationship of p63 and CD44 expressions with age and ultraviolet irradiation in the process of skin aging.Methods Full-thickness skin samples were obtained from the normal skin at different sites,including 44 sun-exposed sites(face,neck,dorsal aspect of the forearm),46 sun-protected sites(abdomen,thigh),and 31 semi-exposed sites(back,flexor aspect of the forearm,upper arm,leg),of 121 volunteers(63 males and 58 females).Of all the volunteers,55 were 10 t0 30 vears of age (young),41 were 31 to 50 years of age(adult),and 25 were 51 to 79 years of age(old).Immunohistochemisty using antibodies to p63 and CD44 was applied to detect the expression of p63 and CD44 in these specimens.Imaging analysis software was utilized to calculate the percentage of p63-expressing cells in basal layer as well as the expression level of p63 and CD44.Results In stratum basale,the percentage of p63-expressing cells decreased with age(r=-0.218,P＜0.05),but showed no significant difference among sun-exposed skin,semi-exposed skin and sun-protected skin(P＞0.05).The correlation between the expression intensity of epidermal p63 and age was negative in sun-exposed sites(r=-0.389,P=0.010),positive in sun-protected sites(r=0.341,P＜0.05).but non-significant in semi-exposed sites(P＞0.05).With regard to the expression of epidermal p63,no significant difference was observed among sun-exposed sites,sun-protected sites and semi-exposed sites in volunteers aged from 10 to 30 years.while in those aged from 31 to 50 years and from 51 to 79 years.sun-protected sites was significantly higher than semi-exposed sites followed by sun-exposed sites(all P＜0.05).There was a weak correlation between the expression intensity of epidermal CD44 and age(r=-0.083,P＜0.05),but no significant difierence was noted among the three kinds of sites(P＞0.05).Conclusions In stratum basale,the percentage of p63.expressing cells and the expression intensity Of CD44 negatively correlate with age,but have no correlation with ultraviolet irradiation.In individuals aged from 31 to 50 years and from 51 t0 79 vears.the expression of epidermal p63 decreases with the accumulation of sun-exposure.
ABSTRACT
Objective: To investigate the role of apoptosis-inducing factor (AIF) in TAp63γ-induced apoptosis. Methods: Plasmid pcDNA3. 1-TAp63γ was transfected into human esophageal squamous cancer EC9706 cells for 24 h. Apoptosis of EC9706 cells was detected by observing DNA fragmentation. The apoptotic rate was measured by flow cytometry. The translocation of AIF in EC9706 cells was studied with Mitochondrial/Cytosol/Nuclear extraction method. The plasmid that expressed hairpin RNA specific for AIF were constructed using chemosynthesis and gene recombination technology, and the interfering efficacy of AIF shRNA was detected by western blotting. After siRNA interference the apoptotic rate of EC9706 cells transfected with pcDNA3. 1-TAp63γ was analyzed by flow cytometry. Results: Agarose gel electrophoresis revealed that DNA ladder, a typical feature of apoptosis, was observed in the pcDNA3. 1-TAp63γ plasmid-transfected cells 24 later, but not in the pcDNA3. 1 plasmid-transfected cells. The apoptotic rate was 13.64% and 1. 37% after EC9707 cells were transfected with pcDNA3. 1-TAp63γ and pcDNA3. 1 plasmid, respectively. Signals of AIF-immunostaining were detected in the nucleus and cytosol proteins of pcDNA3. 1-TAp63γ transfection group, but not in pcDNA3. 1 transfection group. The plasmid expressing shRNA specific for AIF was successfully constructed and the interfering efficiency was about 80%. The apoptotic rate in AIF siRNA group was 4.52% at 24 h after transfection with pcDNA3. 1-TAp63γ. Conclusions: TAp63 γ gene expression induces the apoptosis of EC9706 cells. Down-regulation of AIF protein expression decreases the apoptosis induced by TAp63 γ.