| Project/Area Number |
21791562
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Kyushu University |
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Principal Investigator |
INOUE Takafumi Kyushu University, 大学病院, 助教 (70380417)
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| Project Period (FY) |
2009 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| Budget Amount *help |
¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2010: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2009: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 子宮体がん / アポトーシス / 活性酸素種 / senescence / apoptosis / I型、II型子宮体癌 / K-ras / 分子標的治療 |
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| Research Abstract |
Objectives : Increased expression level of p21 induced by p53 is one of the molecular target of premature senescence. Reactive oxygen species (ROS) is involved in this process. In this study, we investigated how ROS is mediated with the cell growth and mechanism of cellular senescence by suppressed ER using the model of NIH3T3 cell transformed by oncogenic K-ras.
Materials and Methods : 1) We established two NIH3T3 cell lines that expressed only oncogenic K-ras (K12V) or co-expressed dominant negative ER (K12VDNER) and analyed the cell growth. 2) Induction of senescence was evaluated by β-gal staining. Intracellular levels of ROS were measured by FACS using Aminophenyl fluorescin (APF). 3) Inhibitor of ROS, N-acetyl -L-cystein (NAC), was added onto the cell lines, and effect against the induction of senescence was evaluated. 4) Several proteins related with ras signaling was evaluated with immunoblots.
Results : 1) Cell growth of K12VDNER cells were markedly suppressed compared to mock or K12V cells accompanied with senescence by the appearance of multi-nucleated flat cells and significant increase of positive β-gal staining. 3) Intracellular levels of ROS were increased about two times in K12VDNER cells accompanying with induction of senescence. 3) In K12VDNER cell, positive β-gal staining cells were decreased and cell growth was recovered by the addition of NAC. 4) Expression level of p21 was increased in K12VDNER cell compared to mock or K12V cell.
Conclusion : Cellular senescence mediated with oncogenic K-ras and ER related with ROS. We suggested the pathway of senescence via ROS because inhibition of ROS rescued the induction of senescence.
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