| Project/Area Number |
21K07921
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 53010:Gastroenterology-related
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| Research Institution | Osaka Metropolitan University (2022-2023)
Osaka City University (2021) |
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Principal Investigator |
HOANG HAI 大阪公立大学, 大学院医学研究科, 研究員 (60623246)
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| Co-Investigator(Kenkyū-buntansha) |
LE THITHANHTHUY 大阪公立大学, 大学院医学研究科, 准教授 (10572175)
河田 則文 大阪公立大学, 大学院医学研究科, 教授 (30271191)
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| Project Period (FY) |
2021-04-01 – 2025-03-31
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| Project Status |
Granted (Fiscal Year 2023)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2023: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2021: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | DNA methylation / CYGB promoter / Liver tumor / Huh-7 / DAC / Liver cancer / Cytoglobin / Promoter / Demethylation / liver tumor / pancreatic tumor |
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| Outline of Research at the Start |
Cytoglobin (CYGB), an oxygen-binding and reactive oxygen species (ROS) scavenging protein, functions as a tumor suppressor and is inactivated in various cancers. This research aims to (i) examine the protective role of Cygb-overexpression in animal models of liver and pancreas carcinogenesis; (ii) investigate the frequency of DNA methylation of the CYGB promoter in malignant tissues from patients with liver or pancreatic cancers; (iii) targeted demethylation of the CYGB promoter to impair tumor growth. These results will provide the potential therapeutic application of anti-cancer therapy.
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| Outline of Annual Research Achievements |
Cytoglobin (CYGB) acts as a tumor suppressor gene. Restoration of CYGB expression was demonstrated in HCC cell lines treated with 1, 3, 5, and 10 uM 5-aza-2′- deoxycytidine (DAC). Interestingly, DAC treatment time- and dose-dependently restored CYGB expression at both mRNA and protein levels in SNU-387, HLE and Huh7, and at mRNA level in HepG2 cells while DAC did not induce CYGB expression in LX-2. Notably, after inducing CYGB expression in SNU-387, removal of DAC resulted in regressing of CYGB expression at both mRNA and protein levels. Next, we aim to elucidate the DAC is able to inhibit the tumor formation in mice injected with HCC cells.
Huh-7 cells were treated with DAC dose escalation from 1 to 10uM. After 3 weeks of DAC treatment, Huh-7 cells were subcutaneously injected into the right flanks in 9 nude mice. Untreated Huh-7 cells were injected into the left flanks as control.
All of 9 mice appeared big tumors in the left flanks by mean volume of 3570 mm3 while small tumors (mean 61 mm3) were observed in the right flanks injected with Huh-7 + DAC.
Therefore, demethylation of CYGB gene promoter by DAC may lead to cancer regression.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
I have worked at the other institution for 6 months from April to September 2023.
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| Strategy for Future Research Activity |
1- Analyze the tumor formed by Huh-7 cell injection: HE, IF staining.
2- In vitro, liver cancer cells will be investigated CCK8, migration assay, wound healing assay, over expressed CYGB.
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