Molecular cloning of genes expressed downstream the p51 pathway in melanocytes and keratinocytes

黑素细胞和角质形成细胞中 p51 通路下游表达基因的分子克隆

• 批准号: 14570816
• 负责人: YAMASHITA Toshiharu
• 金额: $ 1.86万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570816/
• 关键词: p51(p63); Apoptosis; Transcriptional regulation; Keratinocyte; Melanocyte; Differential display; Micro array; cDNA; CDNA

One of the p53 family members, p51(p63,p73L or Ket) has been reported to be essential for formation of cutaneous organ and adnexal tissues. By using recombinant adenoviruses that express one of the p53 family proteins, suppression of growth and induction of apoptosis were studied in melanoma cell lines. We found that among p53 family proteins, p51 induced apoptosis most significantly than p53 or p73 in melanoma cells. SK-mel-118 melanoma cells infected with p51-expressing adenovirus(Ad-p51) showed fragmented cellular DNA and activated caspase-3 characteristic of apoptosis. We prepared RNA from Ad-LacZ(control)-and Ad-p51-infected SK-mel-118 cells and compared amount of RNA by the methods of differential-display(DD)-PCR and DNA microarray. As a result, we identified more than twenty cellular genes that were clearly induced by p51. Inducibility of the candidate genes were examined in primary fibroblasts, keratinocytes and melanocytes by RT-PCR. Among them, three genes were induced in keratinocytes but not in fibroblasts or melanocytes. From these results, it is suggested that multiple genes including our candidates are induced by p51 and may have some roles in the process of cutaneous formation.

p51（p63、p73L 或 Ket）是 p53 家族成员之一，据报道对于皮肤器官和附件组织的形成至关重要。通过使用表达 p53 家族蛋白之一的重组腺病毒，在黑色素瘤细胞系中研究了生长抑制和细胞凋亡诱导。我们发现，在 p53 家族蛋白中，p51 在黑色素瘤细胞中比 p53 或 p73 诱导细胞凋亡最显着。感染表达p51的腺病毒(Ad-p51)的SK-mel-118黑色素瘤细胞显示出细胞DNA碎片和激活的凋亡特征caspase-3。我们从Ad-LacZ（对照）和Ad-p51感染的SK-mel-118细胞中制备RNA，并通过差异展示（DD）-PCR和DNA微阵列的方法比较RNA的量。结果，我们鉴定了 20 多个明显由 p51 诱导的细胞基因。通过 RT-PCR 检查原代成纤维细胞、角质形成细胞和黑素细胞中候选基因的诱导能力。其中，三个基因在角质形成细胞中被诱导，但在成纤维细胞或黑素细胞中未被诱导。从这些结果来看，包括我们的候选基因在内的多个基因都是由 p51 诱导的，并且可能在皮肤形成过程中发挥一些作用。

项目成果

Sasaki Y et al.: "The p53 family member genes are involved in the notch signal pathway"J Biol Chem. 277・1. 719-724 (2002)

Sasaki Y等：“p53家族成员基因参与Notch信号通路”J Biol Chem.277·1(2002)。

Sasaki Y, Mita H, Toyota M, Ishida S, Morimoto I, Yamashita T, Tanaka T, Imai K, Nakamura Y, Tokino T: "Identification of the interleukin-4 receptora gene as a direct target for p73."Cancer Res. 63. 8145-8152 (2003)

Sasaki Y、Mita H、Toyota M、Ishida S、Morimoto I、Yamashita T、Tanaka T、Imai K、Nakamura Y、Tokino T：“白细胞介素 4 受体基因作为 p73 直接靶标的鉴定。”Cancer Res。

Sasaki Y, Yamashita T et al.: "Identification of the interleukin-4 receptorα gene as a direct target for p73."Cancer Res. 63. 8145-8152 (2003)

Sasaki Y、Yamashita T 等人：“白细胞介素 4 受体 α 基因作为 p73 直接靶标的鉴定。”Cancer Res. 63. 8145-8152 (2003)

Sasaki Y, Ymashita T et al.: "The p53 family member genes are involved in the notch signal pathway."J Biol Chem. 277. 719-724 (2002)

Sasaki Y、Ymashita T 等人：“p53 家族成员基因参与 notch 信号通路。”J Biol Chem。

Takahashi M, Sato T, Segawa T, Lu Y, Sato Y, lyama S, Yamada Y, Fukuda J, Fukaura J, Takahashi S, Miyanishi K, Yamashita T, Sasaki K, Kogawa K, Hamada H, Kato J, Niitsu Y: "E1B-55K-deleted adenovirus expressing E1A-13S by AFP-enhancer/promoter is capable

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