Functional analysis of a novel nuclear peptide H2RSP and its roles in the regeneration of injured gastrointestinal mucosa

新型核肽H2RSP的功能分析及其在损伤胃肠粘膜再生中的作用

• 批准号: 15590351
• 负责人: ITOH Hiroshi
• 金额: $ 2.18万
• 依托单位: University of Miyazaki (2004)宮崎医科大学 (2003)
• 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 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590351/
• 关键词: HGF; HGF activator inhibitor(HAI); HAI-2-related small peptide(H2RSP); gastrointestinal mucosa; Cell growth; Cell differentiation; in situ hybridization; immunohistochemistry; Gastreiatestinal mucisa; Cell differeatiation; In situ hybridization

Hepatocyte growth factor activator inhibitor type 2(HAI-2)-related small peptide(H2RSP) is a small nuclear peptide recently identified in our laboratory. In this study, we examined the expression and cellular localization of H2RSP in normal, hyperplastic, adenomatous and carcinomatous colorectal tissues, and investigated the possible roles of H2RSP on cellular proliferation. Anti-H2RSP polyclonal antibody was raised against the recombinant human H2RSP. Immunohistochemistry and in situ hybridization were performed using surgical specimens of the patients with various gastrointestinal diseases. In normal gastrointestinal mucosae, H2RSP was immunohistochemically localized in the cytoplasm of the epithelial cells at the crypt and translocated into the nucleus of the surface epithelial cells. Its mRNA was detected mainly in the cells at the crypt by in situ hybridization. The hyperplastic epithelial cells showed almost same staining pattern as normal epithelium, while adenoma cells showed d … More ecreased or mixed nuclear and cytoplasmic stainig patterns. In adenocarcinomas, H2RSP was predominantly located in the cytoplasm of almost all cancer cells with various intensities, and nuclear localization was hardly visible. The level of H2RSP mRNA was apparently decreased in the cancer cells compared with corresponding normal epithelial cells by real-time RT-PCR. When cellular localization of H2RSP was examined in fractionated samples of cultured human colorectal carcinoma cells (DLD-1), H2RSP was gradually translocated from the cytoplasm into the nucleus along with the increased cellular density of DLD-1 cells in vitro. In addition, Chinese hamster ovary(CHO) cells stably transfected with H2RSP cDNA revealed the reduced cell growth in vitro. H2RSP was bound to poly(rG)-coated beads, and did not interact with any nuclear protein, single-stranded or double-stranded DNA in pull-down assay using GST-H2RSP fusion protein. The results suggest that H2RSP might be an RNA-binding protein, and have a potentially important role in the proliferation and/or differentiation of the gastrointestinal epithelial cells by translocating from the cytoplasm into the nucleus. Therefore, the impaired nuclear translocation of H2RSP may contribute to colorectal carcinogenesis. Less

肝细胞生长因子激活物抑制剂2型（HAI-2）相关小肽（H2 RSP）是本实验室新近发现的一种核小肽。在这项研究中，我们研究了H2 RSP的表达和细胞定位在正常，增生，腺瘤和癌性结直肠组织，并探讨H2 RSP对细胞增殖的可能作用。制备了针对重组人H2 RSP的抗H2 RSP多克隆抗体。采用免疫组化和原位杂交技术对各种胃肠道疾病患者的手术标本进行检测。在正常胃肠道粘膜中，H_2RSP定位于腺窝上皮细胞的胞浆，并移位到表面上皮细胞的细胞核中。原位杂交结果显示，其mRNA主要分布在腺窝细胞中。增生上皮细胞与正常上皮细胞染色基本一致，腺瘤细胞染色阴性， ...更多信息 核浆染色增强或混合型。在腺癌中，H2 RSP主要位于几乎所有癌细胞的细胞质中，强度不同，核定位几乎不可见。实时荧光定量RT-PCR结果显示，癌细胞中H2 RSP mRNA的表达水平明显低于相应的正常上皮细胞。在体外培养的人大肠癌细胞（DLD-1）的分级样品中检测H2 RSP的细胞定位时，随着DLD-1细胞密度的增加，H2 RSP逐渐从细胞质移位到细胞核中，沿着。此外，稳定转染H2 RSP cDNA的中国仓鼠卵巢（CHO）细胞显示体外细胞生长减少。H2 RSP与poly（rG）包被的微球结合，并且在使用GST-H2 RSP融合蛋白的pull-down测定中不与任何核蛋白、单链或双链DNA相互作用。结果提示，H2 RSP可能是一种RNA结合蛋白，通过从细胞质转位到细胞核，在胃肠道上皮细胞的增殖和/或分化中发挥重要作用。因此，H2 RSP核转位受损可能参与结直肠癌的发生。少

项目成果

Yamauchi M, Kataoka H, Itoh H, Seguchi T, Hasui Y, et al.: "Hepatocyte growth factor activator inhibitor type 1 (HAI-1) and type 2 (HAI-2) are eypressed by epithelium in kidney and downregulated in renal cell carcinoma"J.Urology. 171. 890-896 (2004)

Yamauchi M、Kataoka H、Itoh H、Seguchi T、Hasui Y 等人：“肝细胞生长因子激活剂抑制剂 1 型 (HAI-1) 和 2 型 (HAI-2) 受到肾脏上皮细胞的抑制，并在肾脏中下调

Kataoka H, Tanaka H, nagaike K, Uchiyama S, Itoh H: "Roles of cancer-stroma interaction in invasive growth of cancer cells"Human Cell. 16. 1-14 (2003)

Kataoka H、Tanaka H、nagaike K、Uchiyama S、Itoh H：“癌症-基质相互作用在癌细胞侵袭性生长中的作用”人类细胞。

Characterization of transcripts generated from mouse hepatocyte growth factor activator inhibitor type 2(HAI-2) and HAI-2-related small peptide(H2RSP) genes.

小鼠肝细胞生长因子激活剂抑制剂 2 型 (HAI-2) 和 HAI-2 相关小肽 (H2RSP) 基因生成的转录本的表征。

• 发表时间: 2003
• 期刊: Biochem.Biophys.Res.Commun. 302
• 作者: Naganuma;S.;Itoh;H.;Uchiyama;S.;Tanaka;H.;Nagaike;K.;Miyata;S.;Uchinokura;S.;Nuki;S.;Akiyama;Y.;Chijiiwa;K.;Kataoka;H.
• 通讯作者: H.

Hepatocyte growth factor activator inhibitor type 1 (HAI-1) and type 2 (HAI-2) are expressed by tubular epithelium in kidney and downregulated in renal cell carcinoma.

肝细胞生长因子激活剂抑制剂 1 型 (HAI-1) 和 2 型 (HAI-2) 由肾小管上皮表达，在肾细胞癌中表达下调。

• 发表时间: 2004
• 期刊: J.Urology 171
• 作者: Yamauchi M;Kataoka H;Itoh H;Seguchi T;Hasui Y;et al.
• 通讯作者: et al.

Characterization of transcripts generated from mouse hepatocyte growth factor activator type2 (HAI-2) and HAI-2 related small peptide (H2RSP) genes

小鼠肝细胞生长因子激活剂 2 型 (HAI-2) 和 HAI-2 相关小肽 (H2RSP) 基因生成的转录本的表征

• 发表时间: 2003
• 期刊: Biochem.Biophys.Res.Commun. 302
• 作者: Naganuma S;Itoh H;Uchiyama S;Tanaka H;Nagaike K;et al.
• 通讯作者: et al.