Ann Dermatol.  2016 Jun;28(3):304-313. 10.5021/ad.2016.28.3.304.

Protective Effect of Topical Vitamin D3 against Photocarcinogenesis in a Murine Model

• Kim JS
• Jung M
• Yoo J
• Choi EH
• Park BC
• Kim MH
• Hong SP

• KMID: 2164637
• DOI: http://doi.org/10.5021/ad.2016.28.3.304

Abstract

BACKGROUND
Although the incidence of non-melanoma skin cancer is increasing, there are no effective practical preventive measures other than avoiding sun exposure.
OBJECTIVE
To elucidate the protective effect of topical application of biologically active vitamin D3 (calcitriol) on skin cancer development caused by exposure to ultraviolet (UV).
METHODS
Groups of hairless mice were topically treated with either calcitriol or vehicle immediately after exposure to UVB and UVA three times weekly for the initial 20 weeks, and without UV exposure in the following 6 weeks. Tumor number was counted and biopsies were done for histopathologic analysis. The changes of cyclobutane pyrimidine dimer (CPD) were evaluated 1 hour and 11 hours after short term of UV exposure and application of calcitriol. For safety evaluation, blood test and body weights were evaluated at 23rd and 25th week.
RESULTS
Total tumor count and number of tumors less than 3 mm in size tended to be fewer in calcitriol group, and tumors more than 3 mm in size showed significantly lower tumor formation rate in calcitriol group. Single application of calcitriol reduced CPD at 1 hour and 11 hours after UV exposure. Histopathologic analysis showed tumors with lower grade malignancy in calcitriol group which suggested a delay in tumor progression. However, serum levels of calcium and phosphate in calcitriol group were above normal range, and weight loss was found.
CONCLUSION
Topical calcitriol may suppress the formation and progression of UV-induced non-melanoma skin cancer by enhancing the repair mechanism of UV damage.

Keyword

Calcitriol; Carcinogenesis; Skin neoplasm; 1,25-dihydroxyvitamin D

MeSH Terms

Animals
Biopsy
Body Weight
Calcitriol
Calcium
Carcinogenesis
Cholecalciferol*
Hematologic Tests
Incidence
Mice
Mice, Hairless
Reference Values
Skin Neoplasms
Solar System
Vitamins*
Weight Loss
Calcitriol
Calcium
Cholecalciferol
Vitamins

Figure

• 

  Fig. 1 Gross morphology of dorsum of mouse of each group at 25th week of experiment. Sham light control (A) showed no tumor formation, while vehicle group (B) and calcitriol group (C) showed tumor formation, with vehicle group showing numerous tumors in contrast to few tumors in calcitriol group.

• 

  Fig. 2 Number of tumors in vehicle and calcitriol group between 21st and 26th week of experiment according to different tumor size. (A) Total tumor count (tumors of all size) was higher in vehicle group, without statistical significance except for 21st week. (B) Tumors of 1~3 mm in size were fewer by 2 to 5 on average in calcitriol group, statistically significant at 21st week however not significant thereafter. (C) Tumors more than 3 mm in size showed lower tumor formation rate in calcitriol group compared to vehicle group, with statistical significance at 25th and 26th week. *p<0.05.

• 

  Fig. 3 Histopathological findings of tumor types according to six categories of tumor progression at 26th week of experiment. Dysplastic papilloma with horn cysts: (A) the overall architecture is similar to papilloma with epidermal proliferation and horn cysts without prominent dermal invasion; (B) however it contains dysplastic cells in basal layer. Keratoacanthoma: (C) relatively infiltrative than dysplastic papilloma and individual keratinization begins to appear, (D) with some anaplastic cells. Well-differentiated squamous cell carcinoma (SCC): (E) more than 75% of the tumor is keratinized, (F) with slight amount of anaplastic cells. Moderately differentiated SCC: (G) between 25% and 75% of the tumor is keratinized, (H) with moderate amount of anaplastic cells. Poorly differentiated SCC: (I) less than 25% of the tumor is keratinized, (J) with relatively abundant anaplastic cells. H&E; A, C, E, G, I: ×100; B, D, F, H, J: ×400. Diff.: differentiated, mod.: moderately.

• 

  Fig. 4 Immunofluorescence detection of cyclobutane pyrimidine dimer (CPD) at 1 and 11 hours after ultraviolet-exposure. (A) Vehicle group, (B) calcitriol group. CPD positive cells (yellow-green fluorescence, white arrows) are relatively fewer in calcitriol group than vehicle group (red fluorescence: propidium iodide staining of nuclei). A, B: ×400.

• 

  Fig. 5 Kidney biopsy of mouse in calcitriol group at 26th week of experiment showing non-specific interstitial nephritis. Dense lymphohistiocytic inflammatory cellular infiltrates in interstitial and tubular area (A) and relatively few infiltrates near glomerulus (B). (A, B) H&E, ×200.

• 

  Fig. 6 Laboratory results of vehicle and calcitriol group at 26th week of experiment. In calcitriol group, significantly increased level of calcium (normal range: 7.1~10.1 mg/dl) (A), phosphate (normal range: 5.7~9.2 mg/dl) (B), potassium (normal range: 5~7.5 mEq/L) (C), and ALT (normal range: 17~77 U/L) (D) could be seen, suggesting a hypercalcemic state. AST: alanine aminotransferase.

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