Significance of transforming growth factors for ocular malignant tumors and their application in gene therapy.

转化生长因子对眼部恶性肿瘤的意义及其在基因治疗中的应用。

• 批准号: 15591873
• 负责人: OHNISHI Yoshitaka
• 金额: $ 2.18万
• 依托单位: Wakayama 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: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591873/
• 关键词: Ocular malignant tumor; transforming growth factor; gene therapy; squamous cell carcinoma; XPC knockout mouse; Sonic hedgehog; cyclopamine; 直早期遺伝子

The purpose of this study is to clarify the role of transforming growth factor (TGF) for ocular malignant tumors and to apply them in gene therapy. TGF signaling is required for eliciting the effects of Sonic hedgehog (Shh). Cyclopamine is a natural compound that is capable of suppression of Smoothened activity which is critical in Shh/Gli signal activation.1. Chemical carcinogen cocktail of 9,10-dimethyl-1,2-benzanthracene and phorbol-12-myristate-13-acetate was topically applied to eyelid of XPC-null mice every two days for 2 to 8 months, and epithelial tumors developed in eyelid. These tumors were treated daily with subcutaneous ingestion of cyclopamine for 7 days. Histology showed that cylopamine treatment suppressed BrdU incorporation and induced apoptosis in the majority of cells in these tumors.2. Squamous cell carcinoma cell line established from human burned-scar-derived SCC was used. The cells were maintained in Dulbecco's MEM. The cells were enhanced by adding recombinant Shh and this effect was abolished by adding cyclopamine. However, the proliferation of SCC cell line was not affected by adding cylopamine in the abscence of recombinant Shh.3. Cell suspension of SCC cell line was implanted subcutaneously by using a injection needle in the back of nude mice. Ten days after implantation, the animals were treated systemic cyclopamine at every two days for three times. The SCC cells express Shh in vivo in tumor developed in nude mice and inhibition of Shh signaling pathway by cyclopamine induced apoptosis and suppressed proliferation in the tumor. Cyclopamine inhibits proliferation and induces apoptosis in epithelial tumor cells in vivo. The hedgehog pathway may be a potential therapeutic target for patients with eyelid tumors.

本研究旨在阐明转化生长因子（TGF）在眼部恶性肿瘤中的作用，并将其应用于基因治疗。TGF信号是引发Sonic hedgehog （Shh）作用所必需的。环巴胺是一种能够抑制Shh/Gli信号激活的Smoothened活性的天然化合物。将9,10-二甲基-1,2-苯并蒽和phorl -12-肉豆酸-13-乙酸盐的化学致癌物混合物每2天局部涂于XPC-null小鼠眼睑，持续2 ~ 8个月，眼睑出现上皮性肿瘤。这些肿瘤每天皮下注射环巴胺治疗7天。组织学结果显示，环巴胺处理可抑制BrdU的掺入，并诱导大部分肿瘤细胞凋亡。采用人烧伤瘢痕源性鳞状细胞癌建立的鳞状细胞细胞系。细胞保存在Dulbecco's MEM中。通过添加重组Shh增强细胞，而添加环巴胺则消除了这种作用。然而，在重组Shh.3的培养基中加入环巴胺对SCC细胞株的增殖没有影响。用注射针在裸鼠背部皮下植入SCC细胞系细胞悬液。植入10 d后，给予全身环巴胺治疗，每2天1次，共3次。裸鼠培养的SCC肿瘤细胞在体内表达Shh，环巴胺抑制Shh信号通路可诱导肿瘤细胞凋亡，抑制肿瘤细胞增殖。环巴胺在体内抑制上皮肿瘤细胞增殖和诱导细胞凋亡。刺猬通路可能是眼睑肿瘤患者的潜在治疗靶点。

项目成果

Sonic hedgehog expression and role in healing corneal epithelium

• DOI: 10.1167/iovs.04-0001
• 发表时间: 2004-08-01
• 期刊: INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
• 影响因子: 4.4
• 作者: Saika, S;Muragaki, Y;Kao, WWY
• 通讯作者: Kao, WWY

Saika S: "Smad3 signaling is required for epithelial-mesenchymal transition of lens epithelium post-injury."Am J Pathol. 164. 651-663 (2004)

Saika S：“损伤后晶状体上皮的上皮-间质转化需要 Smad3 信号传导。”Am J Pathol。

Novel complex GUCY2D mutation in Japanese family with cone-rod dystrophy

• DOI: 10.1167/iovs.03-0315
• 发表时间: 2004-05-01
• 期刊: INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
• 影响因子: 4.4
• 作者: Ito, S;Nakamura, M;Miyake, Y
• 通讯作者: Miyake, Y

Fibroblast growth factor 2: roles of regulation of lens cell proliferation and epithelial-mesenchymal transition in response to injury.

• 发表时间: 2004-07
• 期刊: Molecular vision
• 影响因子: 2.2
• 作者: Takeshi Tanaka;S. Saika;Y. Ohnishi;A. Ooshima;J. Mcavoy;Chia-Yang Liu;Muhamad Azhar;T. Doetschman;W. Kao

Osteopontin: a component of matrix in capsular opacification and subcapsular cataract.

• DOI: 10.1167/iovs.02-0873
• 发表时间: 2003-04
• 期刊: Investigative ophthalmology & visual science
• 影响因子: 4.4
• 作者: S. Saika;T. Miyamoto;I. Ishida;Y. Ohnishi;A. Ooshima