Analysis for machanisms invokved in ultraviolct B-light-induced apoptosis in cpidcrmal cells

紫外B光诱导表皮细胞凋亡的机制分析

• 批准号: 11670856
• 负责人: ARAGANE Yoshinori
• 金额: $ 2.11万
• 依托单位: Kinki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670856/
• 关键词: CD95 ligrand(CD95L); ultraviolet B light(UVB); apoptosis; CD95(Fas; APO-a); β-catenin; melanoma; basal call caracinoma(BCC); 遺伝子導入; Fas; Fasリガンド; 前臨床的遺伝子治療; UVB; CD95

This research project was primarily designed to analyze mechanisms involved in ultraviolet B light (UV)-induced apoptosis of epidermal cells. The theoretical background was 1)UV is regarded as a potent carcinogen of non- melanomatous skin cancers, including squamous ell carcinoma (SCC), basal cell carcinoma (BCC), 2)UV is well known to effectively induce apoptosis of epidermal cells, such as keratinocytes and melanocytesl. Based on these so far available knowledge about apidermal cell apoptosis, we thought if we can use the knowlede regarding apoptosis to manipulate therapics, such as those against cancers. Therefore, to address this concern, we first focused on artificial induction of apoptosis of melanoma cells. We previously could chow that melanoma cells, in advanced stage, vigorously express CD95 ligand (CD95L/APO-1L/FasL). Since almost all types of cells including immunocompetent cells express CD95 (Maeda et al, Br J Dermatol 1998 ; 139 : 198-206), melanoma may counterattackimmun … More e cells by inducing their apoptosis via CD95/CD95L crosslinking. To employ this knowledge, we first were interested whether we could induce apoptosis of CD95L-expressing melanoma cells when we introduce expression of CD95 on melanoma cells. The answer was 'yes', as published by us that ectopic expression of CD95 on CD95L-expressing melanoma led to vigorous apoptosis of those cells. This implies future therapeutic uefulness of this strategy in melanoma thereapy.Although the above study clearly indicates the possible effectiveness of CD95 introduction of CD95L+melanoma cells, we should know when melanoma CD95L is turned on. Therefore, we next analysed using patients' specimens for expression of CD95L consequently, it was found that a melanoma cell, when reaches deeper than 3.5-mm from the outside (corresponding approximately middle dermis), becomes able to express CD95L (Maeda et al, J Dermatol 2001 ; 28 : 499-504). Together, we were able to identify the future feasible strategy to treat melanoma patients by ectopic expression of CD95.Besides melanoma, basal cell carcinoma (BCC) is one of the most frequently encountered dermatological neoplasms worldwide. Since pathegenesis of BCC is not completely clear yet, we next focused about this issue. Therefore, we focused β-catenin, a signaling element of Wnt-signaling pathway and reported to be involved in pathogenesis of colon cancers. To address this issue, we immunostained BCC specimens, revealing that nuclear translocation of β-catenin was observed in approximately 70% of BCC tested, while non of other dermatoses, such as atopic dermatitis, psoriasis, squamous cell carcinoma, gave rise to positive signal in muclei. Because nuclear translocation of this protein is a requisite condition to be involved in carcinogenesis, we concluded that b-catenin is involved in pathogenesis of BCC. Further analysis is required to identify which elements are causal to nuclear translocation of β-catenin. Less

该研究项目主要旨在分析紫外线 B 光 (UV) 诱导表皮细胞凋亡的机制。理论背景是 1) 紫外线被认为是非黑色素瘤皮肤癌的强致癌物，包括鳞状细胞癌 (SCC)、基底细胞癌 (BCC)，2) 众所周知，紫外线可有效诱导表皮细胞（如角质形成细胞和黑素细胞）凋亡。基于迄今为止有关表皮细胞凋亡的知识，我们想是否可以利用有关细胞凋亡的知识来操纵治疗，例如抗癌治疗。因此，为了解决这个问题，我们首先关注人工诱导黑色素瘤细胞的凋亡。我们之前可以得知，晚期黑色素瘤细胞强烈表达 CD95 配体 (CD95L/APO-1L/FasL)。由于几乎所有类型的细胞，包括免疫活性细胞，都表达 CD95（Maeda 等人，Br J Dermatol 1998；139：198-206），黑色素瘤可能通过 CD95/CD95L 交联诱导免疫细胞凋亡来反击免疫细胞。为了利用这一知识，我们首先感兴趣的是，当我们在黑色素瘤细胞上引入 CD95 表达时，是否可以诱导表达 CD95L 的黑色素瘤细胞凋亡。答案是“是”，正如我们所发表的，表达 CD95L 的黑色素瘤上 CD95 的异位表达导致这些细胞剧烈凋亡。这意味着该策略在黑色素瘤治疗中的未来治疗价值。虽然上述研究清楚地表明CD95引入CD95L+黑色素瘤细胞的可能有效性，但我们应该知道黑色素瘤CD95L何时被开启。因此，我们接下来使用患者标本分析CD95L的表达，结果发现，当黑色素瘤细胞到达距外部超过3.5mm的深度(大约相当于真皮中层)时，变得能够表达CD95L(Maeda等人，J Dermatol 2001；28：499-504)。我们共同确定了未来通过 CD95 异位表达治疗黑色素瘤患者的可行策略。除了黑色素瘤之外，基底细胞癌 (BCC) 是全世界最常见的皮肤肿瘤之一。由于BCC的发病机制尚不完全清楚，我们接下来重点关注这个问题。因此，我们重点关注Wnt信号通路的信号元件β-连环蛋白，据报道其参与结肠癌的发病机制。为了解决这个问题，我们对 BCC 标本进行了免疫染色，结果显示在大约 70% 的 BCC 测试中观察到了 β-连环蛋白的核转位，而其他皮肤病，如特应性皮炎、牛皮癣、鳞状细胞癌，则没有在细胞核中产生阳性信号。由于该蛋白的核转位是参与癌发生的必要条件，因此我们得出结论，b-catenin 参与了 BCC 的发病机制。需要进一步分析以确定哪些元素导致 β-连环蛋白的核转位。较少的

项目成果

荒金兆典: "サイトカイン産生の調節機構"日本皮膚科学会雑誌. 110. 963-968 (2000)

Arakane, A.：“细胞因子产生的调节机制”，日本皮肤病学会杂志 110. 963-968 (2000)。

Yamazaki F, Aragane Y, Maeda A, Matsushita K, Ueno K, Yudate T, Kawada A, Tezuka T: "Overactivation of IL-r-induced activator protein-1 in atopic dermatitis."J Dermatol Sci. (in press). (2002)

Yamazaki F、Aragane Y、Maeda A、Matsushita K、Ueno K、Yudate T、Kawada A、Tezuka T：“特应性皮炎中 IL-r 诱导的激活蛋白 1 的过度激活。”J Dermatol Sci。

荒金兆典, ほか7名: "Overactivation of IL-4-induced activator protein-1 in atopic dermatitis"Journal of Dermatological Science. (印刷中).

Chonori Arakane 等 7 人：“特应性皮炎中 IL-4 诱导的激活蛋白 1 的过度激活”《皮肤病学杂志》（正在出版）。

前田 晃, ほか5名: "A case of acral lentiginous melanoma : the correlation between CD95L expression on melanoma cells and apoptosis of tumor infiltrating lymphocutes"Journal of Dermatology. 28. 499-504 (2001)

Akira Maeda 等 5 人：“肢端雀斑样黑色素瘤一例：黑色素瘤细胞上 CD95L 表达与肿瘤浸润淋巴细胞凋亡的相关性”《皮肤病学杂志》28. 499-504 (2001)。

荒金兆典ほか5名: "Inhibition of growth of melanoma cells by CD95 (Fas/APO-1) gene transfer in vivo"Journal of Investigative Dermatology. 115. 1008-1014 (2000)

Chonori Arakane 等 5 人：“体内 CD95 (Fas/APO-1) 基因转移抑制黑色素瘤细胞的生长”《皮肤病学研究杂志》115. 1008-1014 (2000)。