Forkhead box protein M1 (FoxM1) 對粒線體動態平衡影響之探討

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研究生：	陳翌萱 Chen, Yi-Hsuan
論文名稱：	Forkhead box protein M1 (FoxM1) 對粒線體動態平衡影響之探討 The Effect of Forkhead box protein M1 (FoxM1) Overexpression on Mitochondrial Dynamics
指導教授：	張壯榮 Chang, Chuang-Rung
口試委員:	王翊青 I-Ching Wang 兵岳忻 Yueh-Hsin Ping
學位類別：	碩士 Master
系所名稱：	生命科學暨醫學院 - 生物科技研究所 Biotechnology
論文出版年：	2014
畢業學年度：	102
語文別：	英文
論文頁數：	70
中文關鍵詞：	粒線體動態平衡、粒線體功能、腫瘤形成
外文關鍵詞：	mitochondrial functions, tumorigenesis
相關次數：	點閱：122 下載：0
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粒線體是不斷進行動態融合分裂的胞器，越來越多研究指出粒線體動態平衡和調控細胞功能有關，包括: 合成ATP，鈣離子濃度平衡，活性氧分子信號途徑和細胞凋亡等。有研究指出粒線體融合和分裂途徑失去平衡，將會導致癌症的產生和轉移。FoxM1是在癌細胞中大量表現的轉錄因子，且和很多癌症的特徵有關，如細胞增生，癌症轉移，血管新生和抵抗細胞凋零。然而，從來沒有研究探討過粒線體動態平衡和FoxM1之間的關係，粒線體動態平衡在FoxM1調控癌症機制中所扮演的角色至今不清楚，因此我們想要串聯兩者間的關係，釐清FoxM1是否會透過調控粒線體動態平衡來促進癌症的產生。我們的研究發現當FoxM1大量表現在人類增殖表皮子宮頸癌細胞(HeLa)時，會改變粒線體動態的平衡，減少族群中含有斷片狀粒線體細胞的比例，同時提高含有長條狀粒線體的細胞，並會降低粒線體活性氧分子，此外，在氧化壓力下，FoxM1大量表現可以保護粒線體的呼吸作用的功能，正常合成能量供給細胞。因此，我們的結果指出FoxM1表現量影響粒線體動態平衡調控和粒線體功能。

Mitochondria are highly dynamics organelles that are regulated by fission and fusion processes. Mitochondria are responsible for many important functions in cells, such as ATP synthesis, calcium homeostasis, ROS signaling and apoptosis. More and more studies have revealed that mitochondrial dynamics is closely correlated with cellular events. In addition, mitochondrial fusion and fission imbalance has been linked to cancer formation and metastasis. FoxM1 is a transcription factor which is overexpressed in cancer cell and associates with many characteristic features of cancer, such as cell proliferation, metastasis, angiogenesis and apoptosis resistance. The relations between mitochondrial dynamics and FoxM1 have not been explored. We aim to clarify whether FoxM1 plays a role in tumorigenesis through affecting mitochondrial dynamics.
In our study, we found that FoxM1 overexpression triggered adjustment of the balance of mitochondrial fusion and fission. In addition, we found FoxM1 overexpression reduced intracellular and mitochondrial superoxide levels. Furthermore, overexpression of FoxM1 aided the mitochondrial respiration activities under induced oxidative stress conditions. Our results indicated that FoxM1 involves in mitochondrial dynamics and adjustments of mitochondrial activities under stress conditions.

誌謝……….................................................    1
中文摘要……...............................................    3
ABSTRACT…………...........................................    4
CONTENTS…...............................................5
LIST OF FIGURES.........................................8
Abbreviations List......................................9
Chapter 1  Introduction................................11
1  Roles of mitochondria in cells....................11
1.1  Structure of mitochondria.......................11
1.2  Functions of mitochondria.......................11
2  Mitochondrial dynamics............................13
2.1  The Mechanism of mitochondrial fusion...........13
2.2  The Mechanism of mitochondrial fission..........14
2.3  Roles of mitochondrial dynamics.................15
2.4  Mitochondrial dynamics and human disease........16
3  Roles of mitochondrial dynamics in cancer.........17
3.1  Mitochondrial dynamics involves in cell death...17
3.2  Mitochondrial dynamics regulates metabolism.....17
3.3  Mitochondrial dynamics promotes ROS production..18
3.4  Mitochondria dynamics participates in migration and invasion...............................................18
4  Roles of the transcription factor Forkhead box protein M1 (FoxM1) in cancer………………………….....………………………………………………….19
4.1  FoxM1 regulates cell cycle and cell proliferation..........................................19
4.2  FoxM1 participates in DNA damage repair and drug resistance.............................................20
4.3  FoxM1 promotes angiogenesis.....................20
4.4  FoxM1 is involved in metastasis.................21
4.5  FoxM1 promotes cell survival....................21
5  The specific aim of this study....................22
Chapter 2  Materials and methods.......................23
1  Cell culture......................................23
2  Cell transfection.................................23
3  Western blot......................................24
4  Immunofluorescence staining.......................25
5  Measurement of mitochondrial membrane potential...25
6  Measurement of cellular bioenergetics.............26
7  Measurement of reactive oxygen species (ROS)......26
8  Measurement of mitochondrial superoxide (O2•−)....27
Chapter 3  Results.....................................28
1  FoxM1 overexpression altered mitochondrial dynamics...............................................28
2  FoxM1 overexpression and dynamics machinery.......29
2.1  FoxM1 overexpression did not affect mitochondrial dynamic proteins expression levels.....................29
2.2  FoxM1 overexpression did not affect phosphorylation levels of Drp1.........................................29
3  Effect of FoxM1 overexpression on mitochondrial bioenergetics and mitochondrial functions..............30
3.1  FoxM1 overexpression did not affect mitochondrial membrane potential (∆ᴪ) and mitochondrial respiration..30
3.2  FoxM1 overexpression decreased intracellular reactive oxygen species level and mitochondrial superoxide level..................................................31
4  Effect of FoxM1 overexpression on mitochondria under oxidative stress.......................................32
4.1  FoxM1 overexpression decreased intracellular reactive oxygen species level and mitochondrial superoxide level under induced oxidative stress...................33
4.2  FoxM1 overexpression maintain mitochondrial respiration under oxidative stress.....................33
Chapter 4  Conclusion and discussion...................35
1  FoxM1 overexpression altered mitochondrial dynamics but not fusion/fission protein expression levels.......35
2  FoxM1 overexpression affected mitochondrial bioenergetics and mitochondrial functions..............38
3  FoxM1 overexpression affected mitochondrial functions under oxidative stress.................................39
Chapter 5  Perspectives................................41
References.............................................42

                                

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