2000 Fiscal Year Final Research Report Summary
Induction of apoptosis via Fas system and enhancement of the induction in bile duct carcinoma cells
Project/Area Number |
11671276
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Digestive surgery
|
Research Institution | Teikyo University |
Principal Investigator |
MARUNO Kaname Teikyo University, Dept.of Surgery, School of Medicine, Assistant Professor, 医学部, 助教授 (20209696)
|
Project Period (FY) |
1999 – 2000
|
Keywords | apoptosis / Fas / human bile duct carcinoma cell / anti-Fas monoclonal antibody / IFN-γ / caspase / IL-2 |
Research Abstract |
Results 1. Fas expression in bile duct carcinoma (BDC) cells, analyzed by flow cytometry : Fas was expressed at 21.7% in HuCCT1 cells and at 30.9% in HuH28 cells, respectively. 2. Viability of BDC cells treated with anti-Fas monoclonal antibody (mAb), CH-11 which induces apoptosis : CH-11 dose-dependently reduced HuCCT1 and HuH28 cell counts at the concentration range from 0.01 to 1.0 μg/ml by up to 16.4% and 71.7% on day 3, respectively. 3. DNA fragmentation in BDC cells treated with CH-11 or IFN-γ : In HuCCT1 and HuH28 cells treated with CH-11 or IFN-γ, the peak corresponding to that of positive control cells was observed. 4. Morphological changes of BDC cells treated with CH-11 or IFN-γ : The shrinkage of cells, chromatin condensation and nuclear fragmentation were observed as morphological changes characteristic of apoptosis. 5. Inhibitory effect of anti-Fas mAb 4B4, which does not induce apoptosis, on the induction of apotosis by CH-11 : 4B4 inhibited the reduction of cell counts by 41
… More
.7% in HuH28 cells. 6. Enhancement of Fas expression in BDC cells treated with IFN-γ : In HuCCT1 cells, the Fas expression was enhanced by 17.6%. In HuH28 cells, however, IFN-γ did not affect the Fas expression. 7. Effect of pretreatment with IFN-γ on the viability of BDC cells treated with CH-11 : Pretreatment with IFN-γ increased the reduction of HuCCT1 and HuH28 cell counts by 41.2% and 8.2% on day 3, respectively. 8. Effect of pretreatment with IL-2 on the viability of BDC cells treated with CH-11 : In HuCCT1 cells, IL-2 alone did not affect the cell number on day 3. Pretreatment with IL-2 also did not affect the reduction of cell counts. In HuH28 cells, IL-2 alone reduced the cell counts by 11.2% on day 3. Pretreatment with IL-2 also did not affect the reduction of cell counts. 9.Viability of BDC cells treated with INF-γ : INF-γ dose-dependently reduced HuCCT1 and HuH28 cell counts at the concentration range from 0.1 to 100 U/ml by up to 45.1% and 87.8% on day 3, respectively. 10. Caspases in BDC cells treated with CH-11 or INF-γ. Any caspase was not detected in HuCCT1 cells. Caspase-3 was detedeted in HuH28 cells treated with CH-11 for 8 hr. Conclusions 1. Fas was expressed in human BDC cells. 2. Apoptosis was induced by anti-Fas mAb and IFN-γ in human BDC cells. 3. Induction of apoptosis by CH-11 was inhibited by 4B4 in human BDC cells. 4. The expression of Fas and the induction of apoptosis was enhanced by IFN-γ in human BDC cells. 5. Caspase-3 was activated in BDC cells treated with anti-Fas mAb. Less
|