| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2005: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2004: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2003: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥700,000 (Direct Cost: ¥700,000)
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| Research Abstract |
The primer set was designed to make an adequate caspase-3 cDNA by polymerase chain reaction (PCR). The caspase-3 cDNA was made by PCR using DNA derived from a healthy individual. Adenovirus vector for caspase-3 was made using Adeno-X^<TM> Expression System (Clontech Laboratories, Inc.). Western blotting showed that all of six gastric cancer cell lines examined but one (KATO-III) expressed much caspase-3 protein. In KATO-3 cells, a faint of caspase-3 protein was detected. Consequently, KATO-III cells were used for caspase-3 gene transfection. The parent KATO-III cells and the caspase-3 gene-transfected cells (KATO-III/cas3) were treated with 5-FU and cisplatin at the concentrations of 0.1, 1, and 10 microgram/ml for 48 hrs. The inhibition rates were measured by the MTT assay. KATO-III/cas3 cells were significantly inhibited, compared to KATO-III cells at any concentrations of 5-FU and cisplatin. These results indicated the caspase-3 gene transfection might be a useful anti-cancer treatment as the combination with chemotherapy. However, the adenovirus vector decreased gradually and disappeared during the culture in HEK293 cells. New adenovirus vectors for caspase-3 gene were tried to make again and again, but in vain. Caspase-3 plasmid could be made successfully. As the second best policy, the hemagglutinating virus of Japan-envelope (HVJ-E) was used to transfect caspase-3 plasmid into gastric cancer cells. However, the cells transfected caspase-3 by this method did not show the significant increment of inhibition rate by chemotherapy, compared to parent cells.
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