| Project/Area Number |
21592057
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Urology
|
|---|
| Research Institution | Nara Medical University |
|---|
Principal Investigator |
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
HIRAO Yoshihiko 奈良県立医科大学, 医学部, 教授 (00133207)
|
|---|
| Project Period (FY) |
2009 – 2011
|
| Project Status |
Completed (Fiscal Year 2011)
|
| Budget Amount *help |
¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2011: ¥260,000 (Direct Cost: ¥200,000、Indirect Cost: ¥60,000)
Fiscal Year 2010: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2009: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
|
|---|
| Keywords | 膀胱癌 / 光力学的診断 / 5-amminolevulinic acid / 尿細胞診 / 遺伝子 |
|---|
| Research Abstract |
The specificity of conventional urine cytology for bladder cancer is high, whereas the sensitivity toward low-grade/low-stage tumors is unsatisfyingly low. In this study, we developed a fluorescence urine cytology system and a protoporphyrinIX(PpIX)-specific fluorescence positive cell detector based on the mechanism of photodynamic reaction of intracellularly accumulated PpIXthat is induced by 5-aminolevulinic acid exposure. Clinical feasibility of fluorescence cytology and the fluorescence positive cell detector was examined by using urine samples from bladder cancer patients and bladder cancer cell lines comparing with conventional cytology. Besides, we investigated significances of genetic alterations of chromosomes 9 and 17 and FGFR3 of tumor cells in urine specimens. Consequently, the PpIX-specific fluorescence positive cell detector, as well as fluorescence cytology, could detect fluorescence positive cells more sensitively in patients with low-grade/low-stage tumors when compared to conventional cytology. Meanwhile, the genetic alterations of shedding tumor cells in urine were highly detected and fluorescence positive tumors included chromosome 9 loss most frequently, p53 loss in high-grade tumors, and FGFR3 mutations in low-stage tumors in incidence order. Peptide nucleic acid real-time PCR clamping or pyrosequencing could detect the genetic alterations even in less shedding cells sensitively. We are currently promoting the system version-up by adopting micro-electro-mechanical systems for cell sorting and a micro-PCR plate for gene analysis to develop a more practical workstation utilizing the above high through-put micro-flow channel systems. The availability for other fluid samples such as ascites, sputum, or blood is also promising.
|
