Change in the Gene Expression Level Observed in the Photodynamically-treated Human Fibroblasts Using cDNA Array

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  • KoH Shunhan
    Department of Biochemistry, Kyorin University School of Medicine
  • YOSHIE Rika
    Department of Trauma and Critical Care Medicine, Kyorin University School of Medicine
  • TSUCHIYA Katsumi
    Department of Biochemistry, Kyorin University School of Medicine
  • SAITO Takashi
    Department of Biology, Kyorin University School of Medicine
  • TAKEICH Toshiaki
    Department of General Medicine, Kyorin University School of Medicine
  • SAKAI Tetsuo
    Department of Biochemistry, Kyorin University School of Medicine
  • TAKAMI Yoshihiro
    Department of Plastic Surgery, Kyorin University School of Medicine
  • HAYASHI Junich
    Department of General Medicine, Kyorin University School of Medicine
  • AIZAWA Katsuo
    Department of Physiology, Tokyo Medical University
  • MURATA Atsuo
    Department of Trauma and Critical Care Medicine, Kyorin University School of Medicine
  • WAKIZAKA Akira
    Department of Biochemistry, Kyorin University School of Medicine

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  • Photodynamic therapy(PDT)によって誘導されるヒト線維芽細胞における遺伝子発現レベル変動のcDNAアレーによる解析
  • Photodynamic therapy PDT ニ ヨッテ ユウドウ サレル ヒト センイ ガ サイボウ ニ オケル イデンシ ハツゲン レベル ヘンドウ ノ cDNA アレー ニ ヨル カイセキ

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Abstract

Development in photosensitizing dyes and laserirradiating equipments have led the photodynamic therapy (PDT) to a clinically useful tool for curing cancer and other diseases in recent years. However, molecular bases of the therapy are not fully understood. We estimated the influence of the therapy on the cells in the expression level in various genes of the cell by using cDNA expression array. Fibroblasts obtained from healthy human skin were administered with mono-Laspartylcholin e6, a photo-sensitizer, at a dose of 10.00 mg/dl for 1 h and irradiated with a diode ion laser beam at 664 nm of wavelength at a total laser energy of 1.00 J/cm^2 cm. The cells were then further incubated at 37℃ for 10 min after the treatment. RNA was extracted from the cells and converted to [^<33>P] labeled cDNA probe by reverse-transcription. The resultant probe was hybridized with a Nylon-membrane CDNA expression array, which was then analyzed for its development using imaging plates and an image scanner. The results showed that the genes related to apoptosis were not increased in their expression level after PDT, except for RARB gene that increased the level after the treatment. While, several other genes related to membrane transports, extracellular communications (mainly growth factors and their receptors), intra-cellular signaling, and transcriptions changed their expression after PDT. This suggested that the treated cells do not need further increase in the level for their apoptosis-related genes but some pathways in cellular regulation system will be changed in the survived cells that escaped from the apoptotic cell death after PDT.

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