| Project/Area Number |
12470431
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Surgical dentistry
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| Research Institution | Hokkaido University |
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Principal Investigator |
TOTSUKA Yasunori Hokkaido Univ., Grad. School of Dent. Med., Prof., 大学院・歯学研究科, 教授 (00109456)
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| Co-Investigator(Kenkyū-buntansha) |
YASUDA Motoaki Hokkaido Univ., Grad. School of Dent. Med., Asso. Prof., 大学院・歯学研究科, 助教授 (90239765)
HIGASHINO Fumihiro Hokkaido Univ., Grad. School of Dent. Med., Inst., 大学院・歯学研究科, 助手 (50301891)
SHINDOH Masanobu Hokkaido Univ., Grad. School of Dent. Med.,Asso. Prof., 大学院・歯学研究科, 助教授 (20162802)
KOBAYASHI Masanobu Inst. Genet. Med., Hokkaido Univ.,Asso. Prof., 遺伝子病制御研究所, 助教授 (80241321)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 2001: ¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 2000: ¥7,600,000 (Direct Cost: ¥7,600,000)
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| Keywords | E1AF / p21 / p53 / cell cycle regulation / 細胞周期調節 |
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| Research Abstract |
E1AF is a member of the ets-oncogene family transcription factor, and E1AF was shown to bind to the promoter region of MMPs that induce transcription from multiple MMP genes. We demonstrated that Ad type 12 E1A is able to suppress the invasion ability of a cancer cell line by inactivating E1AF expression. Ad12E1A expressing oral squamous cell carcinoma cell line was established. Northern blotting indicated that the expression of E1AF mRNA decreased in transfected cells. Reduced expression of MMPs was identified both in mRNA levels and in protein levels. Reduced transcriptional activity of MMP gene was confirmed by CAT assay. These results suggest that Ad12E1A downregulates E1AF which, in turn, restrains invasion ability of human cancer cells by way of downregulaton of MMP expression.
Ca9.22 cells that manifest low levels of E1AF and MMP-1 and -9 expression were transfected with metallothionein inducible E1AF expression vector. MMP-9 and p21_<waf1/cipl> protein were synergistically increased in ZnCl stimulated MT9.22 cells, and MMP-9 and p21 protein were expressed in the same cells when E1AF was induced in MT9.22 cells.
These results imply that invasive growth of tumor cells occur in static state of cell cycle and E1AF play key role of this phenomenon.
Luciferase assays, using different amounts of E1AF and p53 expression vectors along with p2 1 promoter luciferase reporter plasmids, demonstrate an increase in the activity of p21. Colony formation assay indicates that E1AF is capable of tumor suppression similar to p53. These results, therefore, suggest that E1AF is able to interact synergistically with p53 ; in consequence, this interaction can influence p21 leading to an enhancement of its activity and eventually, tumor suppression.
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