Elucidation of a potential role of poly-ADP-ribosylation in the carcinogenesis or development of human colorectal carcinoma and its application for the diagnosis or trteatment of human colorectal carcinoma

阐明聚-ADP-核糖基化在人结直肠癌发生或发展中的潜在作用及其在人结直肠癌诊断或治疗中的应用

• 批准号: 18591456
• 负责人: NABEYA Yoshihiro
• 金额: $ 2.04万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591456/
• 关键词: colorectal cancer; adenosine 5' -diphosphate (ADP)-ribosylation; protein; surgery; ADP-リポシル化

Objective: Poly-adenosine 5'-diphosphate (ADP)-ribosylation, catalyzed by poly (ADP-ribose) polymerase (PARP), transfers poly-ADP-ribose moieties from nicotinamide adenine dinucleotide (NAD) to an acceptor protein. It is an important post-translational modification of nuclear proteins and is involved in various biological processes including malignant transformation. Thus, the aim of this study was to evaluate poly-ADP-ribosylation and to compare the expression of PARP in human colorectal carcinomas (CRCs) and adjacent non-tumorous mucosae.Methods: Tumorous and adjacent non-tumorous tissues were obtained from surgically removed colorectums of 15 patients with CRC, and then were homogenized into cytosolic and membranous fractions. Each homogenate was assayed for ADP-ribosylation with [adenylate-^<32> P] -NAD. The radiolabeled proteins were separated by electrophoresis and autoradiography. The ADP-ribosylated protein was partially purified by anion exchange column chromatography, and analyzed by time-of-flight mass spectrometry (TOF-MS) and immunoprecipitation. Expression of PARP was also evaluated by Western blotting.Results: Several proteins were ADP-ribosylated in human CRC tissues. Notably, the remarkable radiolabeling of a 113-kDa protein was found in the cytosolic fractions from CRCs, and greater than that in adjacent non-tumorous mucosae in 13/15 subjects (87 %). Inhibition assays revealed that the reaction was enzymatic poly-ADP-ribosylation. By TOF-MS and immunoprecipitation, the 113-kDa protein was found to be tumor rejection antigen, glycoprotein (gp) 96. The expression of PARP was significantly greater in CRCs than in adjacent non-tumorous mucosae, while the expression of gp96 in CRCs and normal mucosae was identical.Conclusion: Poly-ADP-ribosylation of gp 96 and PARP expression were increased in human CRCs compared with those in adjacent non-tumorous mucosae, and it may be implicated in the carcinogenesis or development of human CRC.

目的：聚腺苷5 '-二磷酸（ADP）-核糖基化由聚（ADP-核糖）聚合酶（PARP）催化，将聚ADP-核糖部分从烟酰胺腺嘌呤二核苷酸（NAD）转移到受体蛋白。它是一种重要的核蛋白翻译后修饰，参与包括恶性转化在内的多种生物学过程。因此，本研究的目的是评估聚ADP-核糖基化和比较PARP在人结直肠癌（CRCs）和邻近的非肿瘤mucosae.Methods的表达：肿瘤和邻近的非肿瘤组织，从手术切除的结直肠癌患者15例，然后被匀浆成细胞溶质和膜组分。用[腺苷酸-β P] -NAD测定每个匀浆的ADP-核糖基化<32>。通过电泳和放射自显影分离放射性标记的蛋白质。用阴离子交换柱层析法对ADP核糖基化蛋白进行部分纯化，并通过飞行时间质谱（TOF-MS）和免疫沉淀法进行分析。结果：在结直肠癌组织中，有几种蛋白被ADP核糖基化。值得注意的是，在CRC的胞质组分中发现了113-kDa蛋白质的显著放射性标记，并且在13/15例受试者（87%）中大于邻近非肿瘤粘膜中的放射性标记。抑制试验表明，该反应是酶促聚ADP核糖基化。通过飞行时间质谱和免疫沉淀，113 kDa的蛋白被发现是肿瘤排斥抗原，糖蛋白（gp）96。结直肠癌组织中PARP的表达明显高于癌旁组织，而gp 96的表达与癌旁组织无明显差异。结论：结直肠癌组织中gp 96的多聚ADP核糖基化和PARP的表达高于癌旁组织，可能与结直肠癌的发生发展有关。

项目成果

Enhanced poly-(adenosine 5'-diphosphate)-ribosylation of glycoprotein 96 in human colorectal carcinoma

人结直肠癌中糖蛋白 96 的聚（腺苷 5-二磷酸）核糖基化增强

• 发表时间: 2006
• 作者: Nabeya;Y.;Inoue;M.;Yahiro;K.;Morinaga;N.;Sakamoto;A.;Noda;M. and Ochiai;T.
• 通讯作者: T.

Enhanced poly-(adenosine 5'-diphosphate)-ribosylation of glycoprotein 96 in human colorectal carcinoma.

人结直肠癌中糖蛋白 96 的聚（腺苷 5-二磷酸）核糖基化增强。

• 发表时间: 2006
• 作者: Nabeya;Y.;Inoue;M.;et. al. (other 5 authors)
• 通讯作者: et. al. (other 5 authors)

Identification of a protein which can receive poly-(adenosine 5'-diphosphate)-ribosylation in human colorectal carcinoma

人结直肠癌中可接受聚（腺苷5-二磷酸）-核糖基化的蛋白质的鉴定

• 发表时间: 2006
• 作者: Inoue;M.;Nabeya;Y.;Yahiro;K;Akai;T.;Miyazaki;S.;Sakamoto;A.;Noda;M. and Ochiai;T.
• 通讯作者: T.

Enhanced poly- (adenosine 5'-diphosphate) -ribosylation of glycoprotein 96 in human colorectal carcinoma

人结直肠癌中糖蛋白 96 的聚（腺苷 5-二磷酸）-核糖基化增强

• 发表时间: 2006
• 期刊: Abstract DVD of 20th World Congress of International Society for Digestive Surgery 305
• 作者: Nabeya;Y.;Inoue;M.;Yahiro;K;Morinaga;N.;Sakamoto;A.;Noda;M. and Ochiai;T.
• 通讯作者: T.

ヒト大腸癌においてpoly-ADP-ribosylationを受ける蛋白の同定

人类结直肠癌中进行聚 ADP 核糖基化的蛋白质的鉴定

• 发表时间: 2006
• 作者: 井上雅仁;鍋谷圭宏;ほか6名
• 通讯作者: ほか6名