Detection of unique gene in Hypertrophic scar (Keloid) fibroblast by mRNA differential display strangy.

通过 mRNA 差异检测肥厚性疤痕（瘢痕疙瘩）成纤维细胞中的独特基因，结果显示奇怪。

• 批准号: 07671632
• 负责人: MORIGUCHI Takahiko
• 金额: $ 1.6万
• 依托单位: KAWASAKI MEDICAL SCHOOL
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671632/
• 关键词: differential display; hypertrophic scar; Keloid; fibroblasts; Differential Display

Hypertrophic scar (Keloid) can form as a result of deep dermal injury, such as extensive thermal injury, and are characterized by an accumulation of the extracellular matrix in dermis. Lesional skin fibroblasts have been belived to play a crucial role in the disease development. Indeed, abnormal expresions of several extracellular matrix gene in lesional fibroblasts have been observed, however, the alternation mechinery of its function is still unclear. To address molecular mechanisms of change in fibroblasts, we employed a mRNA of differential display which is a novel gene subtraction strategy. Total RNA of cultured dermal fibroblasts derived from involved and uninvolved lesions of Hypertrophic scar patients (n=3) were extracted. Poly-A RNAs were converted into cDNA,amplified by a polymerase chain reaction using 20 sets of primers, then analyzed on a sequencing gel.37 altered clones were detected in involved fibroblasts as compared to uninvolve fibroblasts. 4 clones of them were then sequenced.To analyze details of these genes, full-length cloning and tissue in situ hybridization will be pursued. This approach may lead us to identify gene (s) associated with the nature of the disease.

增生性瘢痕（瘢痕疙瘩）可由于深层真皮损伤（例如广泛的热损伤）而形成，并且其特征在于细胞外基质在真皮中的积累。皮损皮肤成纤维细胞被认为在疾病的发展中起着至关重要的作用。事实上，在病变成纤维细胞中已经观察到多种细胞外基质基因的异常表达，但其功能改变的机制尚不清楚。为了解决成纤维细胞变化的分子机制，我们采用了一种新的基因消减策略，即mRNA差异显示。提取增生性瘢痕患者（n=3）受累和未受累病变的培养真皮成纤维细胞的总RNA。聚-A RNA转化为cDNA，使用20套引物的聚合酶链反应扩增，然后在测序凝胶上进行分析。37个改变的克隆中检测到的成纤维细胞相比，未涉及成纤维细胞。对其中的4个克隆进行了测序，并进行了全长克隆和组织原位杂交分析。这种方法可以帮助我们识别与疾病性质相关的基因。

项目成果

Moriguchi, Takahiko: "Prevention of hypertrophic scar and contracture" J.J.Acute Medicine. 20. 106-107 (1996)

森口隆彦：“预防增生性疤痕和挛缩”J.J.Acute Medicine。

森口隆彦: "創傷治癒.TEXT形成外科学" 南山堂, 11 (1996)

森口贵彦：“伤口愈合.TEXT整形外科” Nanzando，11 (1996)

森口隆彦: "重度皮膚欠損創に対する真皮欠損用グラフト・ペルナックの臨床評価" 薬理と治療. 24. 153-171 (1996)

Takahiko Moriguchi：“真皮缺损移植 Pernac 对严重皮肤缺损伤口的临床评价”药理学和治疗。 24. 153-171 (1996)。

Moriguchi, Takahiko: "The current and future of the biological dressing" JJTOM. 44. 301-306 (1996)

森口隆彦：“生物敷料的现状和未来”JJTOM。

森口隆彦: "創傷被覆材の選択" 川崎医学会誌. 21. 287-296 (1995)

Takahiko Moriguchi：“伤口敷料的选择”川崎医学会杂志 21. 287-296 (1995)。