Research for esophageal carcinogenesis and proliferation, by characterization of cancer stem cells and analysis of the inactivation mechanism for suppressor genes in normal esophageal epithelial cells.

通过癌症干细胞的表征和正常食管上皮细胞抑制基因失活机制的分析来研究食管癌发生和增殖。

• 批准号: 17390363
• 负责人: SHIMADA Yutaka
• 金额: $ 9.79万
• 依托单位: Hyogo College of Medicine (2006)Kyoto University (2005)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 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/grant/KAKENHI-PROJECT-17390363/
• 关键词: Cancer stem cell; Normal esophageal epithelial cell; p75NTR; ABCG2; Hedgeho; Gli-1; NGF; FHIT; Hedgehog; 癌抑制遺伝子

p75NTRp75NTR was expressed in all examined esophageal squamous cell carcinoma (ESCC) and the mean proportion of the p75NTR-positive fraction was 30.1%. The size of p75NTR-positive colonies was larger than that of p75NTR-negative colonies (p<0.0001). Down regulation of p75NTR expression by SiRNA resulted in marked growth inhibition with induction of apoptosis. Our findings suggest that p75NTR was expressed in a small number of cells in ESCC cell lines and p75NTR is necessary for survival and maintenance of ESCC tumors.ABCG2ABCG2 expression was detectable in 61% of patients and the proportion of ABCG2 positive cells was variable (0% to 100%). The presence of ABCG2 positive cells in the tumor was revealed to be an independent prognostic factor along with the extent of the primary tumor and positive lymph node metastasis in multivariate analysis. Our data suggests that ABCG2 plays an underlying role in malignant characteristics of ESCC.GlilCell proliferation and migration were significantly inhibited by Cyclopamine in the cell lines that expressed all Hedgehog (Hh) pathway molecules, while cell proliferation was not affected in the cell lines lacking Shh and Gli-1 expression. SiRNA target for Gli-1 inhibited the cell growth of ESCC cells.FHITNicotine induced methylation of the FHIT gene and attenuated Fhit protein, associated with DNMT3a expression in normal esophageal epithelial cells (HEEC). Furthermore, re-expression of Fhit protein of HEECs was found after cessation of moderate- to long-term exposure to nicotine.NGFNGF overexpression was associated with lymph node metastasis, distant metastasis, higher TNM stage, poorer tumour differentiation and poorer survival of esophageal cancer patients. The motility of HSA/c, one of the ESCC cell lines overexpressing NGF, was significantly decreased by either neutralizing NGF antibody, an inhibitor of TrkA, or NGF-small interfering RNA in transwell migration assay.

p75 NTR在所有食管鳞状细胞癌（ESCC）中均有表达，平均阳性率为30.1%。p75 NTR阳性集落的大小大于p75 NTR阴性集落的大小（p<0.0001）。通过siRNA下调p75 NTR表达导致显著的生长抑制和诱导凋亡。我们的研究结果表明，p75 NTR在ESCC细胞系中表达于少数细胞中，p75 NTR对于ESCC肿瘤的生存和维持是必需的。ABCG 2 ABCG 2在61%的患者中可检测到表达，ABCG 2阳性细胞的比例是可变的（0%至100%）。多因素分析显示，肿瘤中ABCG 2阳性细胞的存在与原发肿瘤的范围和阳性淋巴结转移沿着是独立的预后因素。结果表明，ABCG 2在食管鳞癌的恶性特征中起着重要作用。在表达Hedgehog（Hh）通路分子的细胞系中，环巴胺显著抑制Glil细胞的增殖和迁移，而在Shh和Gli-1表达缺失的细胞系中，细胞增殖不受影响。针对Gli-1的siRNA抑制ESCC细胞的生长，FHIT尼古丁诱导正常食管上皮细胞（HEEC）中FHIT基因甲基化并减弱Fhit蛋白，与DNMT 3a表达相关。NGF-NGF过表达与食管癌患者的淋巴结转移、远处转移、TNM分期、肿瘤分化程度和生存率有关。在transwell迁移实验中，NGF中和抗体、TrkA抑制剂或NGF-小干扰RNA均能显著降低高表达NGF的食管鳞癌细胞系HSA/c的运动能力。

项目成果

An Inducible shRNA Vector against Osteopontin Reduces Metastatic Potential of Human Esophageal Squamous Cell Carcinoma in Vitro and in Vivo

针对骨桥蛋白的诱导型 shRNA 载体可降低人食管鳞状细胞癌的体外和体内转移潜力

• 发表时间: 2006
• 期刊: Clin Cancer Res 12
• 作者: Tetsuo Ito;Yosuke Hashimoto;Eiji Tanaka;Takatsugu Kan;Shigeru Tsunoda;Fumiaki Sato;Motoshige Higashiyama;Tomoyuki Okumura;Yutaka Shimada.;Tsunoda S;Soma T;Mori Y;Okumura T;Soma T;Ito T
• 通讯作者: Ito T

Prognostic significance of fascin overexpression in human esophageal squamous cell carcinoma

• DOI: 10.1158/1078-0432.ccr-04-1378
• 发表时间: 2005-04-01
• 期刊: CLINICAL CANCER RESEARCH
• 影响因子: 11.5
• 作者: Hashimoto, Y;Ito, T;Shimada, Y
• 通讯作者: Shimada, Y

Nicotine induces fragile histidine triad methylation in human esophageal squamous epithelial cells.

尼古丁诱导人食管鳞状上皮细胞中脆弱的组氨酸三联体甲基化。

• 发表时间: 2006
• 期刊: Int J Cancer 119・4
• 作者: Tetsuo Ito;Yosuke Hashimoto;Eiji Tanaka;Takatsugu Kan;Shigeru Tsunoda;Fumiaki Sato;Motoshige Higashiyama;Tomoyuki Okumura;Yutaka Shimada.;Tsunoda S;Soma T
• 通讯作者: Soma T

Significance of nerve growth factor overexpression and its autocrine loop in oesophageal squamous cell carcinoma.

• DOI: 10.1038/sj.bjc.6603255
• 发表时间: 2006-08-07
• 期刊: British journal of cancer
• 影响因子: 8.8

Potential in a single cancer cell to produce heterogeneous morphology, radiosensitivity and gene expression

• DOI: 10.1269/jrr.46.43
• 发表时间: 2005-03-01
• 期刊: JOURNAL OF RADIATION RESEARCH
• 影响因子: 2
• 作者: Ban, S;Ishikawa, K;Imai, T
• 通讯作者: Imai, T