| Project/Area Number |
17590654
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | Kochi University |
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Principal Investigator |
NISHIMORI Isao Kochi University, Kochi Medical School, Department of gastroenterology and Hepatology, lecturer, 医学部附属病院, 講師 (30237747)
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| Co-Investigator(Kenkyū-buntansha) |
TAKEUCHI Tamotsu Kochi University, Kochi Medical School, Department of pathology, Associate professor, 医学部, 助教授 (50226990)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2005: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | carbonic anhydrase / carbonic anhydrase-related protein / colon cancer / small-interfering RNA / microarray / laminin 5 / gastrointestinal stromal cell / lung cancer / 膵癌 |
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| Research Abstract |
The present study investigated molecular function of carbonic anhydrase-related protein (CA-RP) in the cell proliferation of gastrointestinal cancer by using up-and down-regulation techniques of CA-RP expression and then the following results were obtained.
1. Ectopic expression of CA-RP VIII by cDNA transfection in the cultured colon cell (LoVo) enhanced proliferative and invasive abilities as compared to wild-type LoVo cells, which were shown by MTT, colony formation and chemotaxis chamber assays
2..Small-interfering RNA-mediated knockdown of CA-RP VIII revealed significant inhibition in cell proliferation and colony formation of a colon cancer cell line HCT116,which showed high endogenous expression of CA-RP VIII.
3. Ectopic expression of CA-RP XI by cDNA transfection in the cultured gastrointestinal stromal cell (GIST-T1) enhanced proliferative and invasive abilities as compared to wild-type GIST-T1cells, which were shown by MTT and chemotaxis chamber assays
4. Ectopic expression of CA
… More
-RP VIII by cDNA transfection in the cultured lung cancer cell (PC-9) enhanced invasive ability as compared to wild-type PC-9 cells, which was shown by chemotaxis chamber assays
5. Screening analysis for alternatively expressed molecules by ectopic expression of CA-RP VIII in colon cancer cell (LoVo) by using a microarray showed up-and down-regulated expression of several genes. Alternative expression of some molecules were reconfirmed by real-time PCR technique. Among the genes studied, expression of laminin β3 subunit was inhibited by the ectopic expression of CA-RP VIII (0.18 times by array analysis and 0.192 times by real-time PCR). Further, mRNA expression of other subunits of laminin 5,α3 and γ2 subunits, were also inhibited. Decreased expression of laminin 5 has been reported to enhance proliferative and invasive abilities of cancer cells with other tissue types. Taken together, the findings suggest that laminin 5 plays a role in the CA-RP VIII-mediated tumorinogenesis
6. Screening analysis for alternatively expressed molecules by ectopic expression of CA-RP VIII in XI in GIST-T1 by using a microarray showed several genes which was up-regulated (maximum 5.66 times) and down-regulated (minimum 0.09).
These findings strongly indicate that CA-RP (at least VIII and XI) function to enhance proliferative and invasive abilities of tumor cells. Laminin 5 possibly play a role in the CA-RP-mediated tumorinogeneis. Less
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