已知雌激素能夠藉其受體促進各種細胞的生長或抑制凋亡，值得注意的是，高劑量雌激素對於乳癌細胞具有誘導細胞凋亡的現象，然而此功能的詳細作用機制至今仍不甚了解。 KLF10為Krüppel-like factors家族的成員之一，能夠以其锌指的結構與基因啟動子結合並扮演一轉錄調控的角色，且能被雌激素所刺激而增加其表現量；論文中先以染色體免疫沉澱晶片（ChIP-chip）的分析，試圖藉此了解KLF10所可能調控的下游基因，結果顯示其中之一與細胞凋亡有關之基因：BI-1或可解釋KLF10導致細胞凋亡的機制。
本論文陸續以啟動子活性分析（promoter activity assay）、西方墨點染色法（Western blot）及北方墨點染色法（Northern blot）的實驗，先行確認了雌激素/KLF10/BI-1三者間的調控關係，結果顯示雌激素係經由提升細胞中的KLF10蛋白進而抑制BI-1的表現量；其中在啟動子活性分析的實驗經點突變KLF10在BI-1啟動子的位置，或將si-KLF10送入細胞用以抑制KLF10的處理下，皆能夠讓BI-1的啟動子活性恢復到施加雌激素之前的水準。為了深入確認KLF10所造成的影響是否也會反映在BI-1的功能上，實驗針對當BI-1處於環境壓力下會抑制鈣離子從內質網中釋出以避免凋亡的功能做分析，結果顯示KLF10不只抑制了BI-1的表現量，也能夠使細胞質中鈣離濃度上升更甚或是提高細胞死亡率。另外，由於Sp1和KLF 10這兩個轉錄因子所辨識及結合的DNA序列十分相似，再加上曾有文獻指出兩者有競爭同一個啟動子結合位的現象，因此在實驗的最後利用BI-1的啟動子活性分析與電泳遷移率（Electrophoretic Mobility Shift Assay）探討Sp1和KLF 10對BI-1表現上的影響與互動，結果顯示Sp1和KLF 10的確有可能在BI-1的啟動子上競爭同一個結合位，並且對BI-1的表現上有著相反的效果。綜合上述的結果，本論文指出雌激素能夠藉由提升KLF10再降低BI-1表現量的方式，促使細胞質中的鈣離子濃度上升，最後導致細胞的凋亡。

Estrogen stimulates cell growth and inhibits apoptosis through estrogen receptor-mediated mechanisms in many cell types. Remarkably, there is another dimension to estrogen action by which apoptosis is induced in breast cancer cells. While these mechanisms are not yet completely understood, finding the molecules involved has paved the way for the development of a new drug group. Using ChIP-chip, the data have demonstrated that KLF10, a Krüppel-like zinc finger transcription factor, which was induced in response to estrogen, directly modulates the transcription of BI-1 (Bax inhibitor-1; also called TEGT). Eventually, the estrogen induced KLF10 and then suppresses BI-1 transcription. The estrogen/KLF10/BI-1 interrelationship was further confirmed using BI-1 promoter assay、western blot and northern blot. The estrogen-elicited reduction of BI-1 promoter activity was significantly reversed when the KLF10 binding element was mutated to abolish KLF10 binding. A si-KLF10 antisense-oligo nucleotide was also able to restore BI-1 promoter activity to its pre-estrogen-treatment level. BI-1 is known to regulate stress via the endoplasmic reticulum; in this context down-regulation of BI-1 is able to cause Ca2+ release and trigger an apoptosis pathway in breast cancer. In our study, KLF10 not only suppressed cellular BI-1 expression but also increased the cytosolic Ca2+ concentration, eventually causing apoptotic cell death. Besides, Sp1 have zinger motifs similar to KLF10, and it has been identified that KLF family compete with Sp1 for DNA binding site. To study the effect of KLF10 and Sp1 on BI-1 expression, the BI-1 promoter activity and EMSA assay were used. The experiments demonstrate that Sp1 may compete with KLF10 for specific site on BI-1 promoter, and they have opposite function on BI-1 expression. Based on these results, this thesis suggests the pathway by which estrogen induces apoptosis is possibly through an up-regulation of KLF10 that decreases BI-1 and finally increases the concentration of cytoplasmic calcium.

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