Ann Dermatol.  2016 Apr;28(2):179-185. 10.5021/ad.2016.28.2.179.

Potential Role of S100A8 in Cutaneous Squamous Cell Carcinoma Differentiation

• Shin JM
• Chang IK
• Lee YH
• Yeo MK
• Kim JM
• Sohn KC
• Im M
• Seo YJ
• Kim CD
• Lee JH
• Lee Y

• KMID: 2171374
• DOI: http://doi.org/10.5021/ad.2016.28.2.179

Abstract

BACKGROUND
S100A8 is differentially expressed in various cell types and is associated with a number of malignant disorders. S100A8 may affect tumor biology. However, its role in cutaneous squamous cell carcinoma (SCC) is not well established.
OBJECTIVE
This study aims to investigate the relationship between S100A8 and cutaneous SCC development.
METHODS
We performed immunohistochemical staining to detect S100A8 expression in facial skin specimens of premalignant actinic keratosis (AK), malignant SCC, and normal tissues. In addition, we utilized postconfluence and high calcium-induced differentiation in a culture system model. Furthermore, we constructed a recombinant adenovirus expressing GFP-tagged S100A8 to investigate the role of S100A8 in SCC cell differentiation.
RESULTS
S100A8 was significantly overexpressed in human cutaneous SCC compared to that in normal and AK tissues. S100A8 was gradually upregulated in SCC cells in a post-confluence-induced differentiation model. Overexpression of S100A8 in SCC cells induced by adenoviral transduction led to increased expression levels of differentiation markers, such as loricrin, involucrin, and filaggrin. S100A8 overexpression also increased loricrin and involucrin luciferase activity.
CONCLUSION
S100A8 regulates cutaneous SCC differentiation and induces well-differentiated SCC formation in skin.

Keyword

Actinic keratosis; Differentiation; S100; S100A8; Squamous cell carcinoma

MeSH Terms

Adenoviridae
Antigens, Differentiation
Biology
Carcinoma, Squamous Cell*
Cell Differentiation
Humans
Keratosis, Actinic
Luciferases
Skin
Antigens, Differentiation
Luciferases

Figure

• 

  Fig. 1 S100A8 is highly expressed in cutaneous squamous cell carcinoma. (A) Immunohistochemical staining of S100A8 in normal, actinic keratosis (AK), and cutaneous squamous cell carcinoma (SCC) tissues. Positive staining intensity was calculated using an image analyzer. (B) S100A8 protein was detected by western blotting analysis. Lower panel: relative protein levels, which were calibrated to an internal control (actin). Gr: grade. *p<0.05 compared to AK.

• 

  Fig. 2 Increased S100A8 expression in differentiated SCC12 cells. SCC12 cell differentiation was induced using a post-confluence-induced differentiation model or 8.0 mM calcium for 1, 3, or 5 days. S100A8 expression increased in a differentiation-dependent manner. Involucrin and loricrin were used as SCC12 differentiation markers, and actin was used as an internal control.

• 

  Fig. 3 Effect of S100A8 on SCC12 cell differentiation. (A) SCC12 cells were transduced with adenovirus expressing GFP-tagged S100A8 or GFP (control) and cultured for 3 days. Upper layer: bright field, lower layer: fluorescent field (×100). (B) Expression of differentiation-related proteins was assessed by western blotting analysis. S100A8 overexpression in SCC12 cells induced expression of involucrin, loricrin, and filaggrin. (C) An involucrin or loricrin promoter-luciferase reporter adenovirus was co-transduced with an adenovirus expressing GFP-tagged S100A8 or GFP (control). The cells were harvested after 3 days for a reporter assay. Involucrin and loricrin promoter activities are expressed as percentages of the control±standard deviation. *p<0.05 compared to control.

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