Analysis of mechanism of estrogen action on estrogen receptor cDNA transfected endometrial cancer cells
| Project/Area Number |
12670176
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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 |
Human pathology
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| Research Institution | Kitasato University |
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Principal Investigator |
WATANABE Jun Kitasato University, School of Medicine Assistant Professor, 医学部, 講師 (10201188)
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| Co-Investigator(Kenkyū-buntansha) |
DOBASHI You Kitasato University, School of Medicine Associate Professor, 医学部, 助教授 (90231456)
KURAMOTO Hiroyuki Kitasato University, School of Allied Health Sciences Professor, 医療衛生学部, 教授 (80050491)
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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 |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2001: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | endometrial cancer / estrogen / estrogen receptor / gene trasfection / in vitro / cell probferation |
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| Research Abstract |
1. Purpose: This study was designed to establish estrogen responsive human endometrial cancer cell lines positive for estrogen receptor (ER) by trasfecting ER expression vector, and to investigate a mechanism of estrogen action.
2. Methods: Ishikawa cell line was used for the study. It was originally was positive for ER but it became less responsive to estrogen, as the level of ER was gradually decreased. pSG5-HEGO was used as ER expression vector that contained human ER cDNA, wild type. Superfect method was carried out to transfect the vector into the cells. ER transfected cells were cloned by using Neomycin. The levels of ER expression was measured by immunoblotting or enxyme-linked immunoassay. Analysis of cell proliferation was performed by cell growth curve. Effects of estrogen on regulation of cell cycle regulators were examined by immunostaining and Western blotting.
3. Results: ER level of null Ishikawa cells was very low. Thirty six clones were analyzed and three of them expressed high level of ER. Maximal effect of estrogen on cell proliferation was observed at a concentration of 10nM. An 20% increased of growth was detected on Day 6. Levels of Ki-67, Cyclin A, D1, E was also enhanced by estrogen.
4. Conclusion: Stable endometrial cancer cell lines that express high ER and are responsive to estrogen were established. They could be useful to investigate the molecular mechanism of estrogen action.
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Report
(3 results)
Research Products
(14 results)
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[Publications] Akaboshi, M., Watanabe, J., Fujisawa, T., Hattori, M., Ohno, E. and Kuramoto, H.: "Immunohistochemical expression of cdk2 and Ki-67 in human endometrial carcinoma"J. Jap. Soc. Clin. Cytol.. 40. 121-127 (2001)
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[Publications] Fujisawa, T., Watanabe, J., Akaboshi, M., Ohno, E. and Kuramoto, H.: "Immunohistochemical study on VEGF expression in endometrial carcinoma comparison with p53 expression, angiogenesis, and tumor histologic grade"J., J. Cancer Res. Clin. Oncol.. 127. 668-674 (2001)
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[Publications] Watanabe, J., Nishimura, Y., Kato, N., Kamata, Y., Fujisawa, T., Jobo, T. and Kuramoto, H.: "Expression of cell cycle regulators in endometrial carcinoma"J. Jap. Soci. Gyncol. Oncol.. 20. 17-21 (2002)
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[Publications] Kyushima, N., Watanabe, J., Hata, H., Jobo, T., Kameya, T. and Kuramoto, H.: "Expression of cyclin A in endometrial adenocarcinoma and its correlation with proliferative activity and clinicopathological variables"J. Cancer. Res. Clin. Oncol.. (in press). (2002)
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