Project/Area Number |
21590999
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Respiratory organ internal medicine
|
Research Institution | Kyushu University |
Principal Investigator |
|
Co-Investigator(Kenkyū-buntansha) |
HARADA Taishi 九州大学, 大学病院, 講師 (10380619)
IZUMI Miiru 九州大学, 大学病院, 講師 (60336021)
|
Project Period (FY) |
2009 – 2011
|
Project Status |
Completed (Fiscal Year 2011)
|
Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2011: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2010: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2009: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
|
Keywords | 癌 / ウィルス / 細胞・組織 / がん / ウィルスベクター / 細胞診 / 癌性胸水 / ルシフェラーゼアッセイ / GFP |
Research Abstract |
In this study, the adenoviral vector that was modified in fiber region was used as a cancer cell detection tool. The modified adenovirus has a serotype 3 adenoviral fiber protein on the serotype 5 adenovirus, and is reported to show the high infectivity to various cancer cells in the previous paper. The purpose of this study is to improve the diagnosis accuracy for malignant pleuritis or ascites in cytological examination in combination with the modified adenoviral vector. A fire fly luciferase cDNA or GFP cDNA driven by CMV promoter are recombinated into the modified adenoviral genome using the homologous recombination method. Cancer cells infected with this recombinant modified adenoviral vectors showed a high luciferase activity or green lumination under the fluorescent microscopy. Using a diluted serum including fixed number of cancer cells as an artificial malignant effusion, the recombinant modified adenoviral vector was confirmed to detect only 10 cancer cells after RBC cells we
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re lysed out. For the clinical samples including various benign or malignant pleural effusion and ascites, the sensitivity and specificity in the diagnosis were xx% and xx%, respectively. The diagnosis accuracy with this method is a little better than that in usual cytological examination. However, in the malignant effusion with hematological malignancies like a lymphoma, the sensitivity is relatively low probably due to the lack of receptor protein for adenovirus infection on the surface of these malignant cells. On the other hand, some benign effusion also showed the higher luciferase activity resulting a pseudo-positive specimen. With a GFP expressiong modified adenoviral vector, some contaminated mesothelial cells were also infected with viral vector, and might deteriorate the diagnosis specificity. In conclusion, this method is suggested to be useful for cancer cell detection in the effusion samples in combination with cytological examination. Using a tumor specific promoter instead with CMV promoter, the diagnosis accuracy is suggested to be improved. Less
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