Effect of NB-UVB irradiation on the expression of CCL20 in human HaCaT keratinocytes
Objective To investigate the effects of narrow-band ultraviolet B(NB-UVB)irradiation,a psoriasis treatment,on the mRNA and proteinexpression of C-C motif chemokine ligand 20(CCL20)in human immortalized keratinocytes(HaCaT cells)and to explore the underlying mechanisms.Methods HaCaT cells were divided into two groups after routine culture.The first group was exposed to NB-UVB at doses of 0,100,200,and 400 mJ/cm2,and treated after 3,12,and 24 hours of culture.The second group was pre-treated with different concentrations of pyrrolidine dithiocarbamate ammonium(PDTC)(1.0,12.5,25.0 μmol/L),followed by NB-UVB(400 mJ/cm2)exposure.The expression of CCL20 mRNA in the cell culture lysate was detected using the reverse transcription polymerase chain reaction(RT-PCR)method,the expression of CCL20,nuclear factor κB(NF-κB)p65,and phosphorylated NF-κB p65 proteins in HaCaT cells was detected by Western blot,and the expression of CCL20 protein in the cell culture supernatant was measured by enzyme-linked immunosorbent assay(ELISA).Results After NB-UVB irradiation,the CCL20 mRNA expression level in HaCaT cells and the CCL20 protein expression level in cell culture supernatant increased with the increase of NB-UVB irradiation dose,showing a statistically significant difference(P<0.05).The phosphorylation level of NF-κB p65 in HaCaT cells also increased with the increase of NB-UVB irradiation dose,demonstrating a statistically significant difference(P<0.05).Compared with the group treated solely with NB-UVB,the mRNA and protein expression levels of CCL20,as well as the phosphorylation level of NF-κB p65 in HaCaT cells pre-treated with PDTC,were all reduced to varying degrees(P<0.05),and exhibited a dose-dependent manner,with a more pronounced reduction as the concentration of PDTC increased.Conclusions NB-UVB can regulate the expression and secretion of the chemokine CCL20 in human HaCaT keratinocytes through the activation of the NF-κB signaling pathway.
PsoriasisNB-UVBimmortalized human keratinocytesC-C motif chemokine ligand 20nuclear factor κB
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