Identification of differentiation regulated gene and application to differentiation inducing gene therapy

分化调控基因的鉴定及其在分化诱导基因治疗中的应用

• 批准号: 14571210
• 负责人: SHIRABE Ken
• 金额: $ 2.43万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571210/
• 关键词: Hepatocellular carcinoma; induction of differentiation; spheroid; metastasis-related gene; 転移規定遺伝子

1.Identification of differentiation inducing-related genesMicroarray analysis was performed to examine the gene expression induced by HDAC inhibitor. TSA, and spheroid formation. The expressing OcU3/4 gene was induced and Early Growth Response A gene was suppressed in microarray analysis both TSA and spheroid examination. These results suggested that these genes were associated with induction of differentiation. Furthermore, liver specific function -related gene (C/EBPα), and cell cycle -related gene(cyclin B1. topoisomerase II) were clown regulated in TSA and spheroid group. The transfection of HDAC1 antisense to HCC cell line induced the suppression of cell growth and accelerating the synthesis of albumin. Now, DNA microarray analysis was operating.2.Identification of metastasis and /or invasion-related genes in hepatomaWe established the metastatic cell lines (lung, lymph node and intraperitonial metastasis), derived from human cholangiocarcinoma. As a result of DNA microarray analysis, compared with parental cell line, we examined the dickkopf-1 and IL-1 beta, as the intraperitoneal metastasisrelated genes. The IL-1 bet.a transfected cell line showed the increased rate of intraperitoneal metastasis significantly. Wnt signal inhibitor, dickkopf-1, was resulted to be intraperitoneal metastasis in patients with intrahepatic cholangiocarcinoma.. Focal adhesion kinase(FAK), reported as cell migration regulating gene, was recognized that venous invasion and the expression of FAN was correlative in clinical samples. The suppression of Grb7, positioned the lower pathway of FAR, using siRNA inhibited invasion of cancer cells in invasion assay.

1.分化诱导相关基因的鉴定采用基因芯片技术检测HDAC抑制剂诱导分化相关基因的表达。TSA和球体形成。微阵列分析TSA和球体检测均显示OcU 3/4基因表达被诱导，早期生长反应A基因表达被抑制。这些结果表明，这些基因与诱导分化。此外，肝脏特异性功能相关基因（C/EBPα）和细胞周期相关基因（cyclin B1.拓扑异构酶II）在TSA和球状体组中表达下调。HDAC 1反义核酸转染肝癌细胞后，细胞生长受到抑制，白蛋白合成增加。2.肝癌转移和/或侵袭相关基因的鉴定我们建立了人胆管癌转移细胞系（肺转移、淋巴结转移和腹膜内转移）。通过DNA微阵列分析，与亲本细胞系相比，我们检测了dickkopf-1和IL-1 β作为腹膜内转移相关基因。IL-1bet.a转染的细胞系显示腹膜内转移率显著增加。Wnt信号抑制因子dickkopf-1的表达与肝内胆管癌的腹腔转移密切相关粘着斑激酶（Focal adhesion kinase，FAK）是一种细胞迁移调节基因，临床标本中FAN的表达与静脉浸润密切相关。在侵袭实验中，siRNA抑制位于FAR下游通路的Grb 7可抑制癌细胞的侵袭。

项目成果

Shinji Itoh, et al.: "Role of Expression of Focal Adhesion Kinase on Progression in Hepatocellular Carcinoma"Clinical Cancer Research. April, 15(In press). (2004)

Shinji Itoh 等人：“粘着斑激酶的表达对肝细胞癌进展的作用”临床癌症研究。

Shinji Itoh, et al.: "Role of Expression of Focal Adhesion Kinase on Progression in Ilepatocellular Carcinoma"Clinical Cancer Research. (2004)

Shinji Itoh 等人：“粘着斑激酶表达对肠细胞癌进展的作用”临床癌症研究。