1. Vitamin D3 Produced by Skin Exposure to UVR Inhibits Murine Basal Cell Carcinoma Carcinogenesis 
Anastasia Makarova, Grace Wang, John A. Dolorito, Subheksha KC, Eileen Libove, Ervin H. Epstein 
Journal of Investigative Dermatology 
Volume 137, Issue 12, Pages (December 2017)
DOI: /j.jid
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2. Figure 1
Topical vitamin D3 treatment in a dose-dependent manner delays tumorigenesis in PF mice. (a) Sites of BCC analysis. The distance from the site of application of D3 to the site of the biopsy was approximately 3 cm (left). The distance from the site of application to the vBCCs varied from 0 to 10 cm, depending on their location (right). (b) Topical application of 0.38 μg/d D3 significantly reduced the numbers of μBCCs/mm present in the skin of D-depleted PF mice at age 5 months compared with D-depleted PF mice receiving topical acetone or to normal-D fed mice. Mean ± SD. (c) Topical application of high dose of 3.8 μg/d D3 significantly reduced the numbers of μBCCs/mm present in the skin of D-depleted PF mice at age 4 months compared with D-depleted PF mice receiving topical acetone or to normal D-fed mice. Mean ± SD. (d) Kaplan-Meier analysis of vBCC-free survival showed a significant extension (P = 0.0375, log-rank test) of tumor-free survival in mice treated with (3.8 μg/d) topical D3 compared with the topical vehicle-treated group or the dietary D3 group. (e) Topical D3 (3.8 μg/d) reduced the number of vBCCs at age 5–7 months compared with control vehicle-treated mice, whereas dietary D3 reduced vBCC numbers less robustly, if at all. (f) Representative images showing that topical D3, in particular at the higher dose, dramatically reduced the number and size of μBCCs in skin biopsies taken at age 4 or 5 months. Hematoxylin and eosin. Scale bar = 50 μm. (g) In D-deficient PF mice topical D3 (0.38 and 3.8 μg/d five times per week applied for 6–7 months) increased the circulating D3 and 25(OH)D3 blood levels in D-depleted mice compared with control vehicle-treated mice. D3 at 3.8 μg/d raised blood 25(OH)D3 to levels similar to those in mice fed normal D3/normal minerals diet (TD2018) (D3 dietary group). Data are shown as mean ± SD. 25(OH)D3, 25-hydroxy-vitamin D3; BCC, basal cell carcinoma; PF, Ptch1+/– K14-CreER2 p53fl/fl mice; μBCC, microscopic basal cell carcinoma; vBCC, visible basal cell carcinoma; Sc5d, lathosterol 5-desaturase; SD, standard deviation.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
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3. Figure 2
Topical vitamin D3 treatment in a dose-dependent manner decreases cellular proliferation. Topical D3 (0.38 μg/d) decreases cellular proliferation in vivo in μBCCs as shown by the reduction in Ki67 staining at age 5 months. (a) Percentage of μBCCs with one or more Ki67-positive cells (∗P ≤0.001, compared with the control vehicle-treated group) are shown. Mean ± standard deviation. (b) Representative images of μBCCs. Hematoxylin and eosin. (c) D3 in vitro reduces cell proliferation as measured by WST-1 analysis and (d) reduces Gli1 expression as measured by qPCR. μBCC, microscopic basal cell carcinoma; WST, water-soluble tetrazolium.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
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4. Figure 3
UVR treatment accelerates BCC progression and D3 production in male but not female PF mice. (a) Acute UVR treatment with 350 mJ/cm2 UVR on each of three alternate mornings to mice fed a normal D3/normal minerals diet (TD2018) dramatically increased the levels of D3 in blood obtained 6 hours after the third treatment in female PF mice (P < 0.001), but not in male PF mice (P = 0.08), n = 4 mice/group, mean ± SD. (b) UVR treatment increased the numbers of μBCCs/mm present at age 5 months in male (P = 0.036) but not in female PF mice. n = 15 mice/group, mean ± SD. (c–e) In male PF mice fed a normal D3/normal mineral diet (TD2018) UVR significantly reduced tumor-free (P = 0.047, log-rank test) and overall (P = , log-rank test) survival, n = 15 mice/group, and increased the number of vBCCs (P ≤ 0.05). (f–h) In female PF mice fed a normal D3/normal minerals diet, UVR affected neither tumor-free (P = 0.14, log-rank test) nor overall survival (P = 0.2, log-rank test), n = 15 mice/group, and did not alter the number of vBCCs. BCC, basal cell carcinoma; μBCC, microscopic BCC; NS, not significant; PF, Ptch1+/– K14-CreER2 p53fl/fl mice; SD, standard deviation; vBCC, visible BCC.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
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5. Figure 4
Keratinocyte Sc5d deletion sensitizes female mice to UVR stimulation of BCC carcinogenesis. (a) UVR treatment of PF female mice with keratinocyte-specific Sc5D deletion ingesting a normal D3/normal mineral diet (TD2018) significantly increased the numbers of μBCCs/mm present at age 4 months compared with either UVR-untreated control mice or UVR-treated PF mice with wild-type Sc5D expression. Mean ± SD. (b) Representative images of μBCCs in skin biopsies taken at age 4 months. Hematoxylin and eosin. Scale bar = 50 μm. (c) In PF female Sc5Dk14KO mice ingesting a normal D3/normal mineral diet, UVR treatment causes significant shortening of overall survival compared with PF control mice with wt Sc5D expression (with or without UVR treatment), P ≤ , log-rank test. (d) In PF Sc5Dk14KO female mice ingesting a normal D3/normal mineral diet, average tumor volume of vBCCs increases with the duration of UVR treatment but minimally, if at all, in PF Sc5D wt female mice. ∗P = 0.022, mean ± SD. (e) Model illustrating the complex effect of UVR on human skin carcinogenesis: the incidence of SCCs is proportional to lifetime hours of sunlight, but the incidence of BCCs rises until approximately 30,000 hours of exposure and then plateaus (see Rosso et al. [1996a]). BCC, basal cell carcinoma; μBCC, microscopic BCC; PF, Ptch1+/– K14-CreER2 p53fl/fl mice; PF Sc5d, Ptch1+/– K14-CreER2 p53fl/flSc5dfl/fl mice; Sc5d, lathosterol 5-desaturase; SSC, squamous cell carcinoma; SD, standard deviation; vBCC, visible BCC; wt, wild type.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
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