1. Alterations in Cholesterol Sulfate and its Biosynthetic Enzyme During Multistage Carcinogenesis in Mouse Skin 
Kaoru Kiguchi, John DiGiovanni 
Journal of Investigative Dermatology 
Volume 111, Issue 6, Pages (December 1998)
DOI: /j x
Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
A single topical application of either TPA or CHRY caused an overall increase in CS levels, cholesterol sulfotransferase activity, TG-I activity, and DNA synthesis in mouse epidermis. Effects of TPA and chrysarobin on levels of CS (A) cholesterol sulfotransferase activity (B), epidermal TG-I activity (C), and DNA synthesis (D) following a single application of TPA (3.4 nmol) (•), chrysarobin (220 nmol) (▴), or acetone (▪) in mouse epidermis. A single topical application of either TPA or CHRY caused an initial depression in Ch-ST activity followed by an overall increase that led to significantly elevated values of CS. Following TPA treatment, Ch-ST activity was biphasic, whereas CHRY treatment resulted in a single peak at 72 h. DNA synthesis in the epidermis closely paralleled the time course of Ch-ST activity. An elevation in TG-I activity was seen following treatment with either tumor promoter. Although the peaks of TG-I activity preceded CS accumulation, the time course of TG-I activation corresponded to the pattern of changes in CS level. Data represent the mean ± SD in 3–4 mice per group. Results were obtained in three repeat experiments.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Both FTW and tape-stripping induced epidermal hyperplasia. Response of the skin at various time points after tape-stripping (A) and FTW (B). Periodic acid Schiff–hematoxylin staining for tape-stripping.Arrows indicate the basement membranes. Hematoxylin and eosin staining for FTW.Scale bar: 10 μm. Both FTW and tape-stripping induced epidermal hyperplasia with peaks at 120–160 h and 72 h, respectively. Unlike FTW, the basement membrane remained intact after tape-stripping.
Journal of Investigative Dermatology  , DOI: ( /j x)
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4. Figure 3
Tape-stripping caused an overall increase in Ch-ST activity, TG-I activity, and CS levels. FTW caused an increase in Ch-ST activity and CS levels. Activities of Ch-ST (A), TG-I (B), and CS levels (C) during healing after a FTW or tape-stripping. Mice received a 4 cm full-thickness sagittal wound or were wounded by repeat (70 times) application of masking tape to strip already shaved dorsal skin (seeMaterials and Methods for further details). Activities of Ch-ST and TG-I were measured at various time points during wound healing. ○, FTW;♦, tape-stripping. Ch-ST activity was suppressed until 24 h after a FTW, was induced above control levels after 48 h, and peaked at 96 h. Ch-ST activity was suppressed until 24 h after tape-stripping, was induced above control levels after 48 h, and exhibited a broad peak from 72 to 120 h. After tape-stripping TG-I activity was induced and remained at a high level. Tape-stripping induced higher levels of activity than FTW for both Ch-ST and TG-I. CS levels were also induced after 48 h and 72 h following a tape-stripping and FTW, respectively, and exhibited a broad peak for at least 8 d after wounding. Data represent the mean ± SD in 3–5 mice per group from two separate experiments. Values for TG-I activities during the healing of tape-stripping wounds had a maximum variation of 10%.
Journal of Investigative Dermatology  , DOI: ( /j x)
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5. Figure 4
TPA and high Ca2+ induced Ch-ST and TG-I activities in primary mouse keratinocytes. Changes in activities of Ch-ST (A) and TG-I (B) in primary mouse keratinocytes following a single treatment of TPA (1.6 nM) (•), chrysarobin (1 μm) (▴), or acetone (▪), and in keratinocytes switched to a medium with a high concentration of Ca2+ (1.4 mM) (♦). Treatment with TPA and high Ca2+ led to an elevation in Ch-ST and TG-I activity. There was no change in Ch-ST or TG-I activity in keratinocytes treated with either CHRY or okadaic acid. Data represent the mean ± SD of three cultures for each group from three separate experiments.
Journal of Investigative Dermatology  , DOI: ( /j x)
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6. Figure 5
EGF, KGF, and IGF-1 induced Ch-ST activity and DNA synthesis but not TG-I activity. Changes in activities of Ch-ST TG-I (□) and DNA synthesis (▪), determined by thymidine incorporation in DNA into primary mouse keratinocytes 20 h after a single treatment of either epidermal growth factor, keratinocyte growth factor, or insulin-like growth factor-1 (20 ng per ml). All three growth factors induced Ch-ST activity and this increase was associated with a concomitant increase in DNA synthesis. There was no measurable increase in TG-I activity. Data represent the mean ± SD of three separate cultures. *Significantly higher (Student t test. p < 0.002) than control;**not significantly (p > 0.4) different from control.
Journal of Investigative Dermatology  , DOI: ( /j x)
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7. Figure 6
A single topical application of CS caused an increase in TG-I activity of mouse epidermis. Effects of CS (A), DHEA-S (B), and cholesterol (C) on Ch-ST activity (•) and TG-I activity (□) following a single topical application of 1 μmol of CS, DHEA-S, or cholesterol in mouse epidermis. A single topical application of either CS or DHEA-S caused a depression in Ch-ST activity. The activity had returned to near control levels by 72 h after treatment. Following CS treatment, TG-I activity was induced with a peak at 48 h after the treatment. There was no change in TG-I activity following DHEA-S treatment. There was no change in activity of either Ch-ST or TG-I following cholesterol treatment. Data represent the mean ± SD in four mice per group.
Journal of Investigative Dermatology  , DOI: ( /j x)
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8. Figure 7
Increased thickness of epidermis and induction of scaling following multiple topical application of CS (hematoxylin and eosin staining). One micromole of CS (B), DHEA-S (C), or cholesterol (D) were applied six times (every other day). Twenty-four hours after the last treatment skins were removed for histologic examination. (A) Vehicle-treated (acetone:methanol 4:1 vol/vol).
Journal of Investigative Dermatology  , DOI: ( /j x)
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9. Figure 8
CS levels and Ch-ST activity were dramatically elevated in papillomas and SCC. Level of CS (A) and activities of Ch-ST (B) and TG-I (C) papillomas and SCC generated by initiation-promotion regimens. Ch-ST activity was dramatically elevated in all 10 of the papillomas and in 11 of 12 SCC analyzed; however, TG-I activity in papillomas and SCC was essentially the same as in control epidermis. Data represent the mean ± SD of 10 papillomas and 12 SCC. Results were obtained in four repeat experiments. Tumors were harvested at least 1 wk after the last TPA treatment. CTR, control epidermis. **Not significantly (p > 0.4) different from control.
Journal of Investigative Dermatology  , DOI: ( /j x)
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10. Figure 9
Ch-ST activity was significantly elevated in epidermis of Ha-ras transgenic neonates. Activities of Ch-ST (▪) and TG-I (□) in epidermis of Ha-ras transgenics. Ch-ST activity was significantly elevated in the epidermis of transgenic neonates (the age when transgene expression is highest) compared with Ch-ST levels in the epidermis of nontransgenic littermates of the same age. Ch-ST activity returned to near control levels by 7 d after birth. TG-I activity was essentially the same level or slightly higher then control in neonates and adult mice and did not correlate with Ch-ST activity. Data represent the mean ± SD of five mice per group from two separate experiments. Controls were nontransgenic age-matched littermates. **Not significantly (p > 0.4) different from control.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions