Development of a new photo-aging model system: A quick evaluation method for photo-damage utilizing elastin-promoter/green fluorescence protein (GFP) reporter gene transfected dermal fibroblasts

2001 Fiscal Year Final Research Report Summary

• Project/Area Number: 11557062
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 展開研究
• Research Field: Dermatology
• Research Institution: Hirosaki University
• Principal Investigator: KON Atsushi Hirosaki University School of Medicine, Dermatology, Assistant Professor, 医学部・附属病院, 講師 (60271798)
• Co-Investigator(Kenkyū-buntansha): TAMAI Katsuto Hirosaki University School of Medicine, Dermatology, Associate Professor, 医学部, 助教授 (20236730)
• Project Period (FY): 1999 – 2001
• Keywords: photo-aging / elastin promoter / solr elaslosis / ultraviolet / fibroblast / sunscreen

Research Abstract

For the purpose of prevention of photo-aging such as deep wrinkle formation, a great number of sunscreens against broad-band spectrum of solar ultraviolet (UV) has been produced. However, in vivo evaluation of sunscreens for anti-photoaging requires long time. In the present experiments, a new evaluation system have been developed. Cultured mouse fibroblasts (3T3) were transfected with human-elastin promoter linked to green fluorescence protein (GFP) reporter gene. After exposure to UVB, green fluorescence intensity of GFP was observed. In addition, after injection of the cells into the mouse skin, fluorescence intensity was quantified using spectrofluorometer with fiber optics (the SPEX FluoroMax-3 of Jobin Yvon Inc.). The cultured cells expressed green fluorescence in UVB-dose dependent manner. The fluorescence of injected cells was measurable from skin surface. These results indicate that quantitative measurement of fluorescence intensity of injected cells to the mice skin might be useful for early evaluation of solar elastosis formation and protective effect of sunscreens.

Research Products

• [Publications] Hashimoto I, Kon A, Tamai K, Uitto J: "Diagnostic dilemma of "sporadic" cases of dystrophic epidermolysis bullosa : a new dominant or mitis recessive mutation?"Exp Deimatol. 8. 140-142 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Tamai K, Murai T, Mayama M, Kon A, Nomura K, Sawamura D, Hanada K, Hashimoto I, Shimizu H, Masunaga T, Nishikawa T, Mitsuhashi Y, Ishida-Yamamoto A, Ikeda S, Ogawa H, McGrath JA, Pulkkinen L, Uino J: "Recurrent COL7AI mutations in Japanese patients with dystophic epidermolysis bullosa : Positional effects of premature termination codon mutations on clinical severity"J Invest Deimatol. 112. 991-993 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Sawamura D, Ina S, Itai K, Meng X, Kon A, Tamai K, Hanada K, Hashimoto I: "In vivo gene introduction into keratinocytes using jet injection"Gene Ther. 6. 1785-1787 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Tamai K, Murai T, Mayama M, Kon A, Sawamura D, Nomura K, Hanada K, Hashimoto I, Uitto J: "Recent seudies on DNA-based diagnosis for Japanese case of dystrophic epidermolysis bullosa"Molecular Medicine : Novel Findings of Gene Diagnosis, Regulation of Gene Expression, and Gene Therapy. 53-61 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kon A, Vindevoghel L, Hashimoto I, Uitto J, Mauviel A: "TGF-β/Smad and TNF-a /NF-KB upregulate type VII collagen gene expression additively at the transcriptional level"Molecular Medicine: Novel Findings of Gene Diagnosis, Regulation of Gene Expression, and Gene Therapy. 175-183 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Sawamura D, Xianmin M, Ina s, Kon A, Tamai K, Ohe Y, Hashimoto I: "Expression vector with DNA of bovine papilloma virus 1 for keratinocyte gene therapy"J Dermatol Sci. 23. 111-116 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Hashimoto I., Kon A., Tamai K., Uitto J.: "Diagnostic dilemma of "sporadic" cases of dystrophic epidermolysis bullosa: a new dominant or mitis recessive mutation?"Exp. Dermatol.. 8. 140-142 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Tamai K., Murai T., Mayama M., Kon A., Nomura K., Sawamura D., Hanada K. Hashimoto I., Shimizu H., Masunaga T., Nishikawa T., Mitsuhashi Y., Ishida-Yamamoto A., Ikeda S., Ogawa H., McGrath JA., Pulkkinen .L, Uitto J.: "Recurrent COL7A1 mutations in Japanese patients with dystrophic epidermolysis bullosa-Positional effects of premature termination codon mutations on clinical severity"J. Invest. Dermatol.. 112. 991-993 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Sawamura D., Ina S., Itai K., Meng X., Kon A., Tamai K., Hanada K., Hashimoto I.: "In vivo gene introduction into keratinocytes using jet injection"Gene Ther.. 6. 1785-1787 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Murai T., Mayama M., Kon A., Sawamura D., Nomura K., Hanada K., Hashirnoto I., Uitto J.: "Recent studies on DNA-based diagnosis for Japanese case of dystrophic epidermolysis bullosa"Molecular Medicine: Novel Findings of Gene Diagnosis, Regulation of Gene Expression, and Gene Therapy. 53-61 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kon A., Vindevoghel L., Hashimoto I., Uitto J., Mauviel A.: "TGF-/β/Smad and TNF- α/NF-κB upregulate type VII collagen gene expression additively at the transcriptional level"Molecular Medicine: Novel Findings o Gene Diagnosis, Regulation of Gene Expression, and Gene Therapy. 175-183 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Sawamura D., Xianmin M., Ina S., Kon A., Tamai K., Ohe Y., Hashimoto I.: "Expression vector with DNA of bovine papilloma virus 1 for keratinocyte gene therapy"J. Dermatol. Sci.. 23. 111-116 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kon A., Sawamura D., Itai K., Tamai K., Hashimoto I.: "Gene therapy of dystrophic epidermolysis bullosa by introduction of type VII collagen gone to keratinocytes"New direction for cellular and organ transplantation. 175-183 (2000)
  • Description: 「研究成果報告書概要(欧文)」より