Aberrant expression of sphingosine kinase and its pathological significance of malignant tumors and neuronal degenerative diseases

鞘氨醇激酶的异常表达及其在恶性肿瘤和神经退行性疾病中的病理意义

• 批准号: 18590526
• 负责人: MURATE Takashi
• 金额: $ 2.57万
• 依托单位: Nagoya 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-18590526/
• 关键词: sphingosine kinase 1; quantitative RT-PCR; myelodysplastic syndrome; acute leukemia; sphingolipid metabolites; Elecrophysiology; GDNF; MEN2 type tumor; sphingosine kinase 1; MEN2 type tumor; myelodysplastic syndromes; RET signal; promoter anal

1.Sphingolipid metabolic enzymes gene expression of myelodysplastic syndrome (MDS) and acute leukemia was measured by quantitative RT-PCR method. It was shown that sphingosine kinase 1 (SPHK1) was increased in high risk MDS and acute leukemia and that neutral sphingomyelinase 2 (NSMase2) was decreased in bone marrow samples from high risk MDS and acute leukemia Using established leukemia cell lines, further analysis was performed. It was revealed that SPHK1 message, SPHK1 protein and SPHK enzyme activity were well correlated with IC50 of anti-cancer drug, daunorubicin (DA) . Sphingolipid metabolites were measured by LC/MS-MS (liquid column chromatography followed by tandem mass spectrometry) . DA treatment modulated each sphingolipid metabolite level. There was a remarkable heterogeneity of IC50 of DA as well as SPHK expression level. With low dose of DA, DA-sensitive cell lines showed remarkable increase of ceramides as well as decrease of sphingosine 1-phosphate (S1P) compared with DA-resistant cell lines. The ratio of ceramide/S1P was changed greatly with IC50 dose of DA. Our results showed, for the first time, that sphingolipid rheostat model proposed by Spiegel, et. al. can be applied in hematological malignancies.2.Using a GDNF (glial cell line derived neurotrophic factor) -responsive neuroblastoma cell line, TGW, we showed that SPHK1 was increased with GDNF and that the response was mediated through the GDNF receptor, RET. Mutated REF gene was recognized as the pathogenesis of MEN2 type tumors. We also proved that SPHK1 gene expression was increased in MEN2 type tumors and that increased SPHK1 gene expression was involved in its oncogenesis.

1.采用定量RT-PCR方法检测骨髓增生异常综合征（MDS）和急性白血病患者鞘脂代谢酶基因表达。结果表明，在高危MDS和急性白血病中，SPHK 1表达增高，而NSMase 2表达降低。结果表明，SPHK 1的信息、蛋白质和酶活性与抗癌药物柔红霉素（DA）的IC 50值具有良好的相关性。通过LC/MS-MS（液相柱色谱法，然后串联质谱法）测定鞘脂代谢物。DA处理调节每个鞘脂代谢物水平。DA的IC_（50）和SPHK的表达水平存在显著的异质性。在低剂量DA作用下，DA敏感细胞系神经酰胺含量显著增加，而1-磷酸鞘氨醇（S1 P）含量显著降低。神经酰胺/S1 P比值随DA的IC_（50）剂量变化较大。2.使用GDNF（胶质细胞系衍生的神经营养因子）反应性神经母细胞瘤细胞系TGW，我们发现GDNF增加SPHK 1，并且这种反应是通过GDNF受体RET介导的。REF基因突变被认为是MEN 2型肿瘤的发病机制。我们还证实SPHK 1基因表达在MEN 2型肿瘤中增加，并且SPHK 1基因表达增加参与其肿瘤发生。

项目成果

A case of coagulation fator V deficiency caused by compound hetero-zygous mutations in the factor V gene

V因子基因复合杂合突变致凝血因子V缺乏1例

• 发表时间: 2006
• 期刊: Hemophilia 12
• 作者: Yamakage;N.;et. al.
• 通讯作者: et. al.

Increased sphingosine kianse 1 gene expression by glial cell line derived neurotrophic factor (GDNF) and its possible involvement in multiple endocrine neoplasia 2A type (MEN2A) onco-genesis

胶质细胞源性神经营养因子 (GDNF) 增加鞘氨醇 kianse 1 基因表达及其可能参与多发性内分泌肿瘤 2A 型 (MEN2A) 肿瘤发生

• 发表时间: 2007
• 作者: Murakami;M.;et. al.
• 通讯作者: et. al.

Depolarization-induced differentiation of PC12 cells is mediated by phospholipase D2 through the transcription factor CREB pathway

• DOI: 10.1111/j.1471-4159.2007.05085.x
• 发表时间: 2008-03-01
• 期刊: JOURNAL OF NEUROCHEMISTRY
• 影响因子: 4.7
• 作者: Banno, Yoshiko;Nemoto, Satoshi;Nozawa, Yoshinori
• 通讯作者: Nozawa, Yoshinori

Mechanism of vitamin D3-induced transcription of phospholipase D1 in HaCat human keratinocytes

• DOI: 10.1016/j.febslet.2007.03.073
• 发表时间: 2007-05-01
• 期刊: FEBS LETTERS
• 影响因子: 3.5
• 作者: Kikuchi, Ryosuke;Sobue, Sayaka;Murate, Takashi
• 通讯作者: Murate, Takashi

Implications of sphingosine kinase 1 expression level for the cellular sphingolipid rheostat: relevance as a marker for daunorubicin sensitivity of leukemia cells

• DOI: 10.1007/s12185-008-0052-0
• 发表时间: 2008-04-01
• 期刊: INTERNATIONAL JOURNAL OF HEMATOLOGY
• 影响因子: 2.1
• 作者: Sobue, S.;Nemoto, S.;Murate, T.
• 通讯作者: Murate, T.