人類的造血前驅細胞是具有多能性的血液幹細胞，能夠經由骨髓性路徑(myeloid)與淋巴性路徑(lymphoid)分化為不同的血液細胞。這些造血幹細胞如何決定本身的分化方向，是目前學界感興趣的課題。其中，紅血球細胞與巨核細胞(Megakaryocyte)具有許多相似處，因此在分化路徑上被認為是較接近的。有些具多能性的造血細胞株(pluripotent cell lines)能夠在不同的化學物質誘導下進行分化。本研究主要利用兩株人類血癌細胞株，HEL與K-562細胞，在進行藥物處理之後分化方向的進行以及其機制的探討。Protein kinase C(PKC)為一種serine-threonine kinase，目前認為其在細胞的訊息傳遞中扮演重要角色。目前所發現的PKC至少有十二種異構體 。前人研究指出，Phorbol myristate acetate (PMA)能夠使人類造血前驅細胞分化為巨核細胞，但當以PKC抑制劑處理後，細胞便會轉向朝紅血球方向分化。因此目前認為，造血過程(hematopoiesis)決定分化方向的機制可能與PKC各異構體有重要關聯。本研究利用PMA誘導細胞分化為模型，在PMA刺激後，以西方墨點法(Western Blot)觀察不同時間點各PKC 異構體的表達量與活化程度；另一方面，也使用adenyl cyclase活化劑Forskolin和PKD(protein kinase D)抑制劑CID755673前處理細胞，觀察cAMP與PKD對於細胞分化方向的影響，並結合過去文獻，比較各現象的差異，試圖找出可能影響分化方向的關鍵蛋白。

Hematopoietic stem cell (HSC) is derived from blood or bone marrow stem cells that differentiate to a variety of specialized cells through the myeloid or lymphoid pathway. The differentiation commitment of HSC attracts attention throughout the decades. Erythrocytes and megakaryocytes (Mks) differentiated from the same progenitor cells. Many pluripotent hematopoietic cell lines are able to differentiate into megakaryocytes when induced with chemical compounds. In this research, we used human erythroleukemia cell lines, HEL and K-562, and treated with phorbol ester to investigate the mechanism of differentiation. Many studies had point out that protein kinase Cs (PKCs) were important megakaryocytic differentiarion. PKCs are serine/threonine kinases that include at least contain 12 isoforms. It is known that inhibition of PKCs suppresses megakaryocytic differentiation and stimulates erythroid differentiation in HEL cells. However, the role of PKC isoform(s) and its’ signal transduction pathway involved in megakaryocytic differentiation remains unknown. In this study, we used human leukemia cell lines, HEL and K562 cells, treated with phorbol 12-myristate 13-acetate (PMA) or Forskolin (adenyl cyclase activator) and CID755673 (protein kinase D inhibitor) to analyze the expression level and activities of PKC isoforms and the effect of cAMP and PKD in megakaryocytic differentiation.

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