織蛋白去乙醯化酶抑制劑(丙基戊酸)對急性白血病細胞生長、凋亡和分化特性的研究

Characterization of the Histone Deacetylase Inhibitor (Valproic Acid) on the Growth, Apoptosis and Differentiation of Acute Myeloid Leukemia

林秀盆(Hsiu-Pen Lin)

指導教授 ： 劉興璟 博士 ； 林建煌 教授

臺北醫學大學/醫學院/醫學檢驗生物技術學研究所/碩士(2005年)

摘要

論文摘要 論文名稱：織蛋白去乙醯化酶抑制劑(丙基戊酸)對急性白血病細胞生長、凋亡和分化特性的研究 研究所名稱：台北醫學大學生物醫學技術研究所 研究生姓名：林秀盆 畢業時間：93學年度第1學期 指導教授：劉興璟 助理教授 醫學研究所 林建煌 教授 醫學研究所 Epigenetic調控基因表現，對造血作用與白血病的形成有密切關聯。組織蛋白去乙醯化酶是epigenetic 調控的重要成員之一。本研究在探討組織蛋白去乙醯化酶與白血病的增生、細胞分化、細胞凋亡的關係。發現組織蛋白去乙醯化酶抑制劑 (酪酸鈉、酪酸苯丙基戊酸和SAHA)，能有效的抑制人類急性骨髓性白血病細胞株的增生，誘導細胞分化或是促進細胞凋亡作用，酪酸鈉、酪酸苯和SAHA使細胞週期停滯G0/G1期，而丙基戊酸不影響細胞週期。同時發現組織蛋白去乙醯化酶抑制劑，能誘導人類急性骨髓性白血病細胞株走向單核球分化，酪酸鈉能有效誘導CD11c表現，丙基戊酸對誘導CD13表現最好，而四種藥劑都能有效誘導CD14表現，並增加α-Naphthyl Acetate Esterase (NAE) 陽性細胞，但NBT (nitroblue tetrazolium) 還原反應細胞很微量。因此我們針對丙基戊酸，治療人類急性骨髓性白血病單核球細胞株 (THP-1) 的影響，研究發現經由細胞週期蛋白質表現，丙基戊酸誘導 cyclin D1、P21和P27表現， cyclin D1 和 P21誘導骨髓性細胞走向分化。而細胞走向分化可能跟核酸c-Jun和蛋白質表現有關，及 Bcl-2、bid、caspase-9 和 caspase-3 蛋白質表現，誘導骨髓性細胞凋亡作用。凋亡路徑可能是啟動死亡接受器 (death receptor) 與粒線體路徑，而經由 tBid 使兩者路徑有交互聯結，而丙基戊酸活化 JNK 路徑對誘導骨髓性細胞分化作用，可能是重要的關鍵。

關鍵字

織蛋白去乙醯化酶抑制劑 ； 丙基戊酸 ； 凋亡作用 ； 分化作用 ； 死亡接受器 ； JNK 路徑

並列摘要

Abstract Epigenetic control of gene expression plays an important role in hematopoiesis and leukemogenesis. One of the major components in epigenetic regulation of gene expression is histone acetylation.Recent studies have shown that histone deacetylase inhibitors (HDACIs) might be useful for treating hematopoietic malignancies. However the effects of these HDACIs on human acute myeloid leukemia (AML) have not been studied comprehensively. In this study, we examined the effects of several clinically available HDACIs on the proliferation, differentiation and apoptosis of AML in vitro. First HL-60 cells were treated with increasing concentrations of sodium butyrate (SB), a prototypic HDACI, phenylbutyrate (PB), and suberoylanilide hydroxamic acid (SAHA), an HDACI in phase-I clinical trials, valproic acid (VA) , a clinically available agent for neurological disorders.We found that SB, PB, and SAHA were able to arrest cell cycle at G0/G1 phase, but VA did not. SB and VA significantly induced the expression of CD11c and CD13, respectively. CD14 expression was upregulated by all four agents, All four agents induced mild neutrophilic and marked monocytic differentiation evidenced by nitroblue tetrazolium (NBT) tests andα-Naphthyl Acetate Esterase (NAE) staining,respectively. Further elucidation of VA induced apoptosis through both mitochondrial and death receptor pathway.VA induced cyclin D1, p21 and p27 expression, cycle D1 and p21 might relate cell differentiation,so cell cycle did not arrest at G0/G1 phase in 48H.Valproic acid activated JNK pathway might play an important role in monocytic differentiation.

並列關鍵字

Epigenetic ； histone deacetylase inhibitors ； valproic acid ； differentiation ； death receptor ； JNK pathway ； apoptosis

參考文獻

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