| Co-Investigator(Kenkyū-buntansha) |
HIKIZI Kazumasa SRL (Lts) Center of gene laboratory, 遺伝子検査センター, 室長
HIRAKATA Youichi Nagasaki University, Hospital of medicine and Dentistry, Lecturer, 医学部・歯学部附属病院, 講師 (50238341)
NAKAGOE Toru Nagasaki University, Graduate School of Biomedical Sciences, Associate Professor, 大学院・医歯薬学総合研究科, 助教授 (40188917)
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| Budget Amount *help |
¥12,400,000 (Direct Cost: ¥12,400,000)
Fiscal Year 2003: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2002: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2001: ¥7,600,000 (Direct Cost: ¥7,600,000)
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| Research Abstract |
Malignant tumor develops based on accumulation of gene mutations, indicating that it is clinically significant for screening, assessing, and monitoring tumors to detect the mutated genes involved in tumorgenesis. However, it is difficult to gain clinical specimens of the tumor materials including appopriate genomic DNAs, because tuomr is generally and deeply located in the body. Accordingly, we examined the usefulness and paradigm' of gene analysis using the parctical spcecimes from plasma collected from patients with solid tumor, as follows, (1) plasma nucleic acid (NA) was quantitatively and qualitatively assessed, (2) quantification of survivin mRNA and detection of k-ras in plasma NA, (3) quantification of survivin and CEA proteins. by ELISA. First of all, plasama DNA level of patients with solid tumor in stage I, II, III, and IV was 1 8.0, 6.0, and 10.3 ng/ml (median), respectively. On the other hand, plasma NA (mainly RNA) level was 47.6, 49.2, 57.3, and 60.3 pg/ml, respecitvely (healthy individuals, 54.4 pg/ml). These plasma NA was confirmed to be appopriate for gene tests because hosekeeping genes of beta-globin and GAPDH are relevantly detectable using LightCycler Reatl Time PCR method. Then, using such materials, k-ras mutation was detected in 7 (7.58%, colon 12 or 13) out of 80 cases, whereas survivin mRNA was detectnle in 5% (4/80). On the other hand, in respect to the protein level of survivin in plasma in reference to the COV of 130 pg/ml, the detecting rate of the tumor in stage I, II, III, and IV was 30 (6/20), 28 (5/18), 30 (6/20), and 27 (6/22), respectively. This results were not associated with CEA plasma level and were mutually exclusive. Conclusively, problematic issues in gene diagnosis of tumor using plasma remains to be elucidated, but it should be dissolved near future for ideal medicine.
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