Gene expression profiling and corresponding sub-classification of fibroblastic cells within neoplastic and non-neoplastic diseased conditions

肿瘤和非肿瘤疾病状态下成纤维细胞的基因表达谱和相应的亚分类

• 批准号: 17590303
• 负责人: NAKAYAMA Hirofumi
• 金额: $ 2.24万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590303/
• 关键词: Fibroblast; pericryptal fibroblast; subserosal fibroblast; CD34; cytokeratin 8; podoplanin; gene expression profile; magnetic beads; 低分子量サイトケラチン; アルファ平滑筋アクチン; 筋線維芽細胞; 潰瘍; 出血性梗塞; M2A oncofetal antigen

The purpose of this study is gene expression profiling analysis and corresponding sub-classification of fibroblastic cells within neoplastic and non-neoplastic diseased conditions.(1) Immunohistochemical analysis using paraffin-embedded sections and monoclonal antibodies to CD34, vimentin(vim), podoplanin/D2-40(pod), cytokeratin 8(CK8), alpha-smooth muscle actin (ASMA), and high molecular weight caldesmon (HCD) was performed in gastric and colorectal neoplastic and non-neoplastic diseases. Seven kinds of fibroblastic cells were proposed; i) CD34+vim+pod-CKS-ASMA-HCD-cells in normal tissue, ii) CD34-vim+pod+CK8-ASMA-HCD-cells in early phase of cancer invasion to submucosa, acute inflammation and recent localized circulatory disturbance, and post-irradiated tissue, iii) CD34-vim+pod+CK8-ASMA+HCD-cells in desmoplastic stroma of invasive adenocarcinomas and periglandular areas of gastrointestinal adenomas, iv) CD34-vim+pod-CK8-ASMA+HCD-cells in scar tissue, v) CD34-vim+pod+CK8+ASMA+HCD-cel … More ls seen in subserosal tissue involved by neoplastic and non-neoplastic diseases, vi) CD34-vim+pod-CK8-ASMA+HCD+ cells in normal colorectal crypts (pericryptal fibroblast), vii) CD34-vim+pod-CK8-ASMA+HCD- cells in peri-glandular areas of gastrointestinal adenomas.(2) To confirm and/or revise the proposed subtypes of fibroblastic cells, cell isolation (fibroblastic cells, lymphocytes, etc) should be hurried from the fresh normal and diseased tissues, using cell type specific cell surface markers and magnetic beads (magnetic bead method). And then, both serial analysis of gene expression and DNA microarray analysis should be done.(3) In performing the magnetic beads method, to choose good cell surface markers for fibroblasts is most important. Both normal fibroblasts and endothelial cells express CD34 in the cell surface, but epithelial cells, mature lymphocytes and other mesenchymal cells does not; normal fibroblast are detected by CD34 in a fraction which endothelial cells are removed by CD31 antibody.(4) No specific cell surface markers exist for cancer-associated fibroblasts (CA-Fib). Most of the cancer-associated fibroblasts express membrane-truncated matrix metalloproteinase (MT1-MMP) in their cell membrane. Specific antibodies to MT1-MMP are suitable for CA-Fib. Less

本研究的目的是对肿瘤和非肿瘤疾病条件下的成纤维细胞进行基因表达谱分析和相应的亚分类。(1) 使用石蜡包埋切片和 CD34、波形蛋白 (vim)、podoplanin/D2-40(pod)、细胞角蛋白 8(CK8)、单克隆抗体进行免疫组织化学分析。 α-平滑肌肌动蛋白（ASMA）和高分子量卡尔德斯蒙（HCD）在胃和结直肠肿瘤和非肿瘤疾病中进行。提出了七种成纤维细胞； i) 正常组织中的 CD34+vim+pod-CKS-ASMA-HCD-细胞，ii) 癌症侵入粘膜下层、急性炎症和近期局部循环障碍以及照射后组织的早期阶段的 CD34-vim+pod+CK8-ASMA-HCD-细胞，iii) 癌症中的 CD34-vim+pod+CK8-ASMA+HCD-细胞 侵袭性腺癌和胃肠道腺瘤腺周区域的促纤维增生基质，iv) 疤痕组织中的 CD34-vim+pod-CK8-ASMA+HCD-细胞，v) CD34-vim+pod+CK8+ASMA+HCD-cel … 更多见于肿瘤和非肿瘤疾病涉及的浆膜下组织， vi) 正常结直肠隐窝（隐窝周围成纤维细胞）中的 CD34-vim+pod-CK8-ASMA+HCD+ 细胞，vii) 胃肠道腺瘤腺周围区域中的 CD34-vim+pod-CK8-ASMA+HCD- 细胞。(2) 为了确认和/或修订所提出的成纤维细胞亚型，细胞分离 应使用细胞类型特异性细胞表面标记物和磁珠（磁珠法）从新鲜的正常和病变组织中快速提取（成纤维细胞、淋巴细胞等）。然后，进行基因表达的系列分析和DNA微阵列分析。(3)在磁珠法中，选择良好的成纤维细胞表面标志物是最重要的。正常成纤维细胞和内皮细胞细胞表面均表达CD34，但上皮细胞、成熟淋巴细胞等间质细胞不表达；在CD31抗体去除内皮细胞的部分中，正常成纤维细胞可被CD34检测到。(4)不存在癌症相关成纤维细胞(CA-Fib)的特异性细胞表面标记。大多数癌症相关成纤维细胞在其细胞膜上表达膜截短的基质金属蛋白酶（MT1-MMP）。 MT1-MMP 特异性抗体适用于 CA-Fib。较少的

项目成果

Lack of pericryptal fibroblastic cells adjacent to intestinal epithelial metaplastic gastric glands

肠上皮化生胃腺附近缺乏隐窝周围成纤维细胞

• 发表时间: 2006
• 期刊: Histopathology 48・5
• 作者: Oda W;Mistrikova J;Stancekova M;Dutia BM;Nash AA;Takahata H;Jin Z;Oka T;Hayashi.;Nakayama H
• 通讯作者: Nakayama H

Cytokeratin-positive subserosal myofibroblasts in gastroduodenal ulcer; another type of myofibroblasts

胃十二指肠溃疡中细胞角蛋白阳性浆膜下肌成纤维细胞；

• DOI: 10.14670/hh-21.697
• 发表时间: 2006-07-01
• 期刊: HISTOLOGY AND HISTOPATHOLOGY
• 影响因子: 2
• 作者: Guo, L;Kuroda, N;Enzan, H
• 通讯作者: Enzan, H

Cytokeratin-positive subserosal myofibroblasts in gastrointestinal ulcer; anthoer type of myofibroblasts.

胃肠道溃疡中细胞角蛋白阳性浆膜下肌成纤维细胞；

• 发表时间: 2006
• 期刊: Histology and Histopathology 21(7)
• 作者: Kondo E;Miyake T;Shibata M;Kimura T;Iwagaki H;Nakamura S;Tanaka T;Ohara N;Ichimura K;Oka T;Yanai H;Shibasaki F;Yoshino T;Guo L
• 通讯作者: Guo L

Lack of pericryptal fibroblastic cells adjacent to intestinal epithelial metaplasic gastric glands.

肠上皮化生胃腺附近缺乏隐窝周围成纤维细胞。

• 发表时间: 2006
• 期刊: Histopathology 48(5)
• 作者: Oda W;Mistrikova J;Stancekova M;Dutia BM;Nash AA;Takahata H;Jin Z;Oka T;Hayashi;Nakayama H
• 通讯作者: Nakayama H

Molecular characteristics of diferentiated type gastric carcinomawith distinct mucin phenotype ; Li-cadherin is associated with intestinal phenotype

具有独特粘蛋白表型的分化型胃癌的分子特征；

• 发表时间: 2006
• 期刊: Pathology International in press
• 作者: Shimakage;M.;Inoue;N.;Oshima;K.;Oka;T.;Tambe;Y.;Yasui;K.;Matsumoto;K.;Yutsudo;M.;Inoue;H.;Motoshima J
• 通讯作者: Motoshima J