Induction of ultraviolet induced melanoma in the reconstructed skin using DNA repair deficient melanocytes

1.我们成功地培养了缺乏DNA修复能力的A型着色性干皮病患者的黑素细胞。与正常人黑素细胞相比，较低剂量的紫外线（UV）B照射更易诱导着色性干皮病黑素细胞发生凋亡，而UVA照射对着色性干皮病黑素细胞的诱导作用无明显差异。2.从遗传背景为C57/B16、黄色和白化病突变的XPA基因敲除小鼠中，建立了不同黑素亚型的DNA修复缺陷型黑素细胞。XPA基因敲除白化病小鼠由日本大坂大学Kiyoji Tanaka教授获得，黄、黑小鼠由我们通过回交建立。3.在SCID小鼠背部用正常人黑素细胞、角质形成细胞和成纤维细胞通过Hat & Cap法制备重建皮肤。成功地制作了黑色的人皮肤，然而，通过组织化学检查，人黑素细胞不在表皮中，而是在真皮中观察到。这表明，黑素细胞存活因子，如SCF（干细胞因子）的添加可能需要维持表皮中的黑素细胞。

1.We succeeded in culturing melanocytes derived from xeroderma pigmentosum type A patients who lacke DNA repair ability. Compared with normal human melanocytes, apoptotic change was easily induced by ultraviolet (UV)B irradiation at lower dose in xeroderma pigmentosum melanocytes, however, there was no difference recognized in case of UVA radiation.2.DNA repair deficient melanocytes with different melanin subspecies were established from XPA gene knockout mice with the genetic background of C57/B16, yellow, and albino mutant. XPA gene knockout albino mice were obtained from Prof. Kiyoji Tanaka in Osaka University and yellow, and black mice were established by us by backcrossing the mice.3.Reconstructed skin was made using normal human melanocytes, keratinocyte, and fibroblasts by Hat & Cap method on the back of SCID mice. Black colored human skin was successfully made, however, by histochemistry examination, human melanocytes were observed not in the epidermis, but in the dermis. This indicates that the addition of melanocyte surviving factor such as SCF (stem cell factor) might be required for maintenance of melanoyctes in the epidermis.