| Co-Investigator(Kenkyū-buntansha) |
KUWABARA Yoshiyuki Nagoya City University, Graduate School of Medical Sciences, Associate Professor, 大学院・医学研究科, 助教授 (90225326)
ISHIGURO Hideyuki Nagoya City University, Graduate School of Medical Sciences, Research Associate, 大学院・医学研究科, 助手 (10363920)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2004: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Research Abstract |
Transfection did siRNA into cell lines TE1 and 2 which TCF expression is high, from among esophageal cancer cell lines TE series, using Oligofectamine. After transfection, we examined the RNA expression after 48 hours because the apparant inhibition was seen most 48 hours later. Moreover, it was 30-40% though the efficiency of transfection was different according to the cell. The expression of TCF4 was measured by using Light Cycler RT-PCR, measured, and decreased in 90% or more at the mRNA level of TCF4 was confirmed compared with the control.
Oligofectamine was used for esophageal cancer cell lines (TE1,2) and colon cancer cell line (SW480,LoVo), siRNA was compared with the transfection doing and the control, and the cell death was detectable in all the cell lines.
A decrease of the number of cells of the inhibition group was confirmed with Giemsa stain by present. However, a significant difference has not been obtained at that time when MTTassay was performed. It was not possible to perform it to the FACS analysis. SiRNA is made again again, and being analyzed now.
RNA was extracted by first using cell line HET1A and 15 kinds of the TE series, and cDNA was made. The microarray of about 30000 genes was done by using the cDNA. It interrupts now though the cluster is analyzed in cell line group from which the cell death is not derivable with cell line group from which the cell death is derivable by siRNA, and the key gene that induced the cell death was scheduled to be identified. As for the result of these microarray, it is scheduled to apply it to the identification of the anti-cancer drug receptivity gene and the development research into new anti-cancer drug.
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