細胞內鈣離子的恆定對許多細胞生理功能的調控上扮演重要的角色，如細胞附著、移動、增生和基因表現。鈣池調控鈣離子流入 (store-operated calcium entry; SOCE) 是在非興奮性細胞中調控鈣離子流入的主要機制。基質交互分子 (stromal interaction molecule 1; STIM1) 是位於細胞內質網膜上的鈣離子感測器，當內質網中的鈣離子濃度下降時，STIM1 會活化並聚集，進而和鈣池調控鈣離子通道蛋白 Orai1 產生交互作用，啟動 SOCE。微管尖端蛋白 (end-binding protein 1; EB1) 是一種位於微管正端的微管結合蛋白，能幫助許多已知的微管正端追蹤蛋白結合到微管上，並能調控微管的動態平衡。在先前的研究中指出微管在 SOCE 活化的過程中及調控 STIM1 在細胞內分佈位置扮演重要的角色。但是目前對於微管調控 STIM1 移動到細胞膜邊的機制尚未明瞭。我的研究目標在於探討 EB1 在 STIM1 活化過程中及鈣離子的流入所扮演的角色。為了研究 EB1 對於 STIM1 在細胞中移動的影響，我使用基因方法在細胞中降低 EB1 的表現。由免疫螢光染色的結果顯示，在 Thapsgargin (Tg) 及 EGF 的刺激下，會促使 STIM1 聚集成團並隨著時間增加其移動細胞膜邊的量，同時其與 EB1 之間的結合率也會增加。但在降低 EB1 表現的細胞中，STIM1 雖然在細胞質中聚集但移動到細胞膜邊的程度減低。而從單細胞鈣離子濃度分析的結果也顯示降低 EB1 的表現會降低 SOCE 的活化。而先前的研究指出，EB1 及組蛋白去乙醯酶 6 (HDAC6) 的表現會影響到微管乙醯化的程度，進而影響到微管的穩定性。同時 HDAC6 的表現也會影響SOCE 的活化。為了研究 HDAC6 是否參與在 EB1 和 STIM1 的交互作用中，我使用 tubastatin A 來抑制 HDAC6 的活性，發現 tubastatin A 雖然能抑制 STIM1 的移動，但並不會對 EB1 和 STIM1 的交互作用造成影響。綜合以上實驗結果，證明 EB1 在子宮頸癌細胞的 STIM1 活化過程中及鈣離子的流入扮演重要的角色，並可能成為未來臨床治療的標靶。

Store-operated calcium entry (SOCE) is the major Ca2+ entry mechanism in non-excitable cells and plays an important role in a wide variety of cellular functions, including adhesion, motility, gene expression and proliferation. Stromal interaction molecule 1 (STIM1) is an endoplasmic reticulum (ER) Ca2+ sensor which is essential for SOCE. End-binding-protein 1 (EB1), a microtubule plus-end binding protein (+TIP), controls microtubule dynamics and interacts with most other known +TIPs to target them to growing microtubule plus-ends. Previous studies indicated that microtubules play a facilitative role in SOCE by optimizing the localization of STIM1. However, the mechanisms of microtubule in STIM1 membrane trafficking remain controversial. My study aims to investigate the roles of microtubule-associated proteins on the STIM1-mediated SOCE Ca2+ signal. In order to study the role of EB1 on STIM1 membrane trafficking, I used genetic approach. Results of immunofluorescent images showed that EB1 knockdown altered STIM1 membrane trafficking. Consistently, the results of single cell intracellular Ca2+ measurement showed that EB1 knockdown inhibited the activation of SOCE. Taken these results together, I suggest that EB1 is necessary for STIM1 membrane trafficking. Previous study showed that acetylated--tubulin levels were reduced in EB1-silencing condition. Moreover, histone deacetylase 6 (HDAC6), a member of the class II histone deacetylase that deacetylates microtubule, can alter the microtubule stability and play a role in STIM1 activation. To investigate whether HDAC6 plays a role in EB1-STIM1 interaction, I used tubastatin A, a potent and selective HDAC6 inhibitor, and found that HDAC6 inhibitor reduced the level of juxta-membrane STIM1, but did not affect the STIM1-EB1 interaction. Taken together, I suggest that end-binding protein 1 is necessary for STIM1 trafficking and is involved in the regulation of SOCE activation in cervical cancer cells. Therefore, EB1 may be a potential therapeutic target for tumor malignant behaviors.

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