| Project/Area Number |
20590566
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Nagoya University |
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Principal Investigator |
MURATE Takashi Nagoya University, 医学部, 教授 (30239537)
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| Co-Investigator(Kenkyū-buntansha) |
KOJIMA Tetsuhito 名古屋大学, 医学部, 教授 (40161913)
TAKAGI Akira 名古屋大学, 医学部, 助教 (30135371)
SUZUKI Motoshi 名古屋大学, 大学院・医学系研究科, 講師 (80236017)
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| Co-Investigator(Renkei-kenkyūsha) |
BANNO Yoshiko 岐阜大学, 医学部, 准教授 (50116852)
NOZAWA Yoshinori 財団法人岐阜県研究開発財団, 岐阜県国際バイオ研究所, 研究所長 (10021362)
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| Project Period (FY) |
2008 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2010: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2009: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2008: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | セラミド / スフィンゴシン1リン酸 / スフィンゴシンキナーゼ1 / スフィンゴシンキナーゼ2 / セラミドキナーゼ / 中性スフィンゴミエリナーゼ2 / スフィンゴシン1リン酸リアーゼ / 転写調節 / ホスホリパーゼD1,ホスホリパーゼD2 / sphingosine 1-phosphate lyase / human lung cancer cell lines / real time RT-PCT / cellular S1P level / promoter analysis / DNA pulldown assay and chromatin immunoprecipitation assay / Sp1 protein / GATA-4 protein / PLD1 transcription / 脂肪分化ならびに癌化 / 転写因子およびプロモーター領域結合部位 / ATRA / 転写抑制機序 / ダウノルビシン / mRNA寿命の延長 / v-SRCガン遺伝子 / 3'-UTR / AUF1 / HuR / タンパク質リン酸化 / PKC alpha |
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| Research Abstract |
The gene expression of human sphingosine 1-phosphate lyase (SPL) catalyzing sphingosine 1-phosphate (S1P) remains to be determined. Among 5 human lung cancer cell lines examined, SPL protein levels paralleled the respective mRNA and enzyme activities. Between H1155 and H1299 cells used for further experiments, lower cellular S1P was observed in H1155 with higher SPL activity compared with H1299 with lower SPL activity. Previous reprots by others have shown that GATA-4 affect SPL transcription in Dictyostelium discoideum. GATA-4 was observed in H1155 but not in other cell lines. Promoter analysis of human SPL revealed that the most important region was located between -136 bp and -88 bp from the first exon, where 2 Sp1 sites exist but no GATA site. Electrophoresis mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP) assay, reporter assay using reporter with mutated binding motif suggest the major role of Sp1 in SPL transcription and no direct binding of GATA-4 with this 5' promoter region. The collive role of GATA-4 wasproved by showing co-immunoprecipitation of Sp1 and GATA-4. Thus, high SPL transcription of H1155 cells was regulated by Sp1 and GATA-4/Sp1 complex formation, both of which bind to Sp1 sites of the 5' -SPL promoter.
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