| 研究実績の概要 |
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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| 今後の研究の推進方策 |
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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