Experimental gene therapy employing DUSP6/MKP-3 for pancreatic cancer

使用 DUSP6/MKP-3 治疗胰腺癌的实验性基因疗法

• 批准号: 13670161
• 负责人: FURUKAWA Toru
• 金额: $ 2.3万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670161/
• 关键词: DUSP6; MKP-3; pancreatic cancer; Tumor suppressor gene; Gene therapy; MAPK; Adenovirus; Apoptosis; LoH; がん抑制遺伝子; 遺伝子治療; アデノウイルス; apoptosis; phosphatase

The loss of heterozygosity at 12q21 and 12q22-q23.1 is frequently found in primary pancreatic cancers ard DUSP6/MKP-3 gene residing in this region at 12q22 lost its expression in the great majority of pancreatic cancer cell lines. The DUSP6/MKP-3 protein is a dual spcificity phosphatase that dephosphorylates the active form of ERK, making a feed back loop to control ERK activity. Gain-of-function mutations of KRAS2 occur ih the great majority of pancreatic cancer cells, and loss of expression of DUSP6/MKP-3 may synergistically promote constitutive activation of ERK and uncontrolled cell growth. To study loss of the feed back pathway and its impact to pancreatic cancer cell growth, the expression of DUSP6/MKP-3 in primary pancreatic cancer tissues was investigated immunohistochemically ; upregulation in mildly as well as severely dysplasticlin situ carcinoma cells and downregulation in invasive carcinoma, especially that of the poorly differentiated type were found. Adenovirus-mediated reintroduction of DUSP6/MKP-3 into cultured pancreatic cancer cells induced strong expression of recombinant DUSP6/MKP-3 and reductin of phosphorylated ERK in a dose-dependent manner based on the multiplicity of infection and resulted in suppression of cell growth. Moreover, analyses by flow cytometry and immunocytochemistry revealed that the exogenous expression of DUSP6/MKP-3 induced apoptosis. In an experimental gene therapy for xenographted human pancreatic cell tumors in nude mice revealed transient retardation of tumor growth. However, dose and administrative ways should be altered for more efficient therapy result. These results shovv that DUSP6 exerts apparent tumor-suppressive effects, and suggest that DUSP6 is a strong candidate tumor suppressor gene at 12q22 locus and one of therapeutic targets of human pancreatic cancer.

原发性胰腺癌中常见12 q21和12 q22-q23.1的杂合性缺失，位于12 q22区域的DUSP 6/MKP-3基因在绝大多数胰腺癌细胞系中表达缺失。DUSP 6/MKP-3蛋白是一种双特异性磷酸酶，其使ERK的活性形式去磷酸化，形成反馈环以控制ERK活性。KRAS 2的功能获得性突变发生在绝大多数胰腺癌细胞中，并且DUSP 6/MKP-3表达的缺失可协同促进ERK的组成性激活和不受控制的细胞生长。为了研究反馈途径的丧失及其对胰腺癌细胞生长的影响，用免疫组织化学方法研究了DUSP 6/MKP-3在原发性胰腺癌组织中的表达;在轻度以及严重的异型增生原位癌细胞中上调，在浸润性癌中下调，腺病毒介导的DUSP 6/MKP-1基因的再导入，3诱导重组DUSP 6/MKP-3的强表达和磷酸化ERK的减少，并导致细胞生长抑制，这是基于感染复数的剂量依赖性方式。流式细胞术和免疫细胞化学分析显示外源性DUSP 6/MKP-3的表达诱导了细胞凋亡。在裸鼠异种移植的人胰腺细胞肿瘤的实验性基因治疗中，显示肿瘤生长的短暂阻滞。然而，应改变剂量和给药方式以获得更有效的治疗结果。这些结果表明DUSP 6具有明显的抑瘤作用，提示DUSP 6是一个位于12 q22位点的强候选抑癌基因，是胰腺癌治疗的靶点之一。

项目成果

Furukawa,T., Sunamura,M., Motoi,F., Matsuno,S., and Horii,A.: "Potential tumor suppressive pathway involving DUSP6/MKP-3 in pancreatic cancer"American Journal of Pathology. in press..

Furukawa,T.、Sunamura,M.、Motoi,F.、Matsuno,S. 和 Horii,A.：“胰腺癌中涉及 DUSP6/MKP-3 的潜在肿瘤抑制途径”美国病理学杂志。