Immune Netw.  2010 Apr;10(2):75-80. 10.4110/in.2010.10.2.75.

The Production IL-21 and VEGF in UVB-irradiated Human Keratinocyte Cell Line, HaCaT

• Kim H
• Kang JS
• Lee WJ

• KMID: 2150662
• DOI: http://doi.org/10.4110/in.2010.10.2.75

Abstract

BACKGROUND: Ultraviolet B (UVB) induces multiple inflammatory and carcinogenic reactions. In skin, UVB induces to secrete several kinds of inflammatory cytokines from keratinocytes and also increases angiogenic process via the modulation of vascular endothelial growth factor (VEGF) production. Interleukin-21 (IL-21) is an inflammatory cytokine and produced by activated T cells. The biologic functions of IL-21 have not yet extensively studied.
METHODS
In the present study, we investigate the production of IL-21 from human keratinocyte cell line, HaCaT and its biological effect after exposure to UVB.
RESULTS
First, we confirmed the IL-21 production and its receptor expression in HaCaT. And then, the change of IL-21 and VEGF production in HaCaT by UVB irradiation was examined. Not only IL-21 but also VEGF production was enhanced by UVB irradiation. Next, to determine relationship of enhanced production of IL-21 and VEGF, we detected VEGF production after neutralization of IL-21. VEGF production was reduced by IL-21 neutralization, which indicates that the IL-21 is involved in the VEGF production.
CONCLUSION
Taken together, our results suggest that IL-21 and VEGF production is enhanced by UVB irradiation in HaCaT. In addition, it seems that IL-21 plays a role in the angiogenic process in skin via the modulation of VEGF production.

Keyword

HaCaT; UVB; IL-21; VEGF

MeSH Terms

Cell Line
Cytokines
Humans
Interleukins
Keratinocytes
Skin
T-Lymphocytes
Vascular Endothelial Growth Factor A
Cytokines
Interleukins
Vascular Endothelial Growth Factor A

Figure

• 

  Figure 1 IL-21 production and its receptor expression in human keratinocyte cell line, HaCaT. (A) HaCaT cells (1×106) were harvested, intracellular IL-21 was measured by flow cytometry as described in Materials and Methods. (B) HaCaT cells (1×106) were harvested, the surface expression of IL-21 receptor was examined by flow cytometry as described in Materials and Methods.

• 

  Figure 2 IL-21 production from HaCaT cells by UVB irradiation. (A) HaCaT cells (7×106) were exposed to 0, 100, 150, 200, 300 J/m2 of UVB and incubated for 6 hrs. After, total RNA was isolated and cDNA was synthesized, RT-PCR for IL-21 was performed. β-actin was used as a loading control. (B) HaCaT cells (3×106) were exposed to 150 J/m2 of UVB and incubated for overnight. After cells were harvested, IL-21 production was measured by intracellular flow cytometry.

• 

  Figure 3 VEGF production from human keratinocyte cell line, HaCaT by UVB. HaCaT cells (3×106) were exposed to 50, 100 J/m2 of UVB and incubated for overnight. Then, culture supernatants were collected, and VEGF concentration was measured by ELISA.

• 

  Figure 4 Inhibition of UVB-induced VEGF production by the treatment of soluble IL-21 receptor. HaCaT cells (3×106) were pre-treated with sIL-21R for 2 hrs before exposure to 100 J/m2 of UVB. After UVB irradiation, cells were incubated for overnight. And then, culture supernatants were collected, and the concentration of VEGF was measured by ELISA.

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