Hispolon 對人類急性前骨髓血癌細胞株(NB4)誘導細胞凋亡之分子機制探討;Molecular Mechanisms of Hispolon Induced Apoptosis in Human Acute Promyelocytic Leukemia NB4 Cell Line.

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Title:	Hispolon 對人類急性前骨髓血癌細胞株(NB4)誘導細胞凋亡之分子機制探討;Molecular Mechanisms of Hispolon Induced Apoptosis in Human Acute Promyelocytic Leukemia NB4 Cell Line.
Authors:	郭婉琳;Wan-Lin Kuo
Contributors:	中國醫藥大學：中國藥學研究所
Keywords:	桑黃;血癌;細胞凋亡;細胞週期;Hispolon;NB4;apoptosis;cell cycle
Date:	2008-06-27
Issue Date:	2009-08-10 18:02:14 (UTC+8)
Abstract:	桑黃 (Phellinu merrillii)為東方人(尤其是中國、日本、南韓)的傳統中藥，目前在抗菌、抗癌、抗纖維化、抗氧化及抗發炎方面皆有相關的研究，且桑黃為已知之高等真菌中抗腫瘤效果最具效果的菌類。其中具有高度抗氧化能力的化合物之一的Hispolon，已有研究指出可促使KB cell(口腔癌細胞)的細胞凋亡，然而Hispolon對人類急性前骨髓血癌細胞株(NB4)產生的細胞凋亡影響仍不明確。本次研究中，主要為探討其對細胞產生的毒性、細胞凋亡及細胞週期的影響，以及凋亡的相關訊息路徑。在細胞毒性方面，藉由Trypan blue染色計數、DAPI染色、DNA斷裂和流式細胞儀分析，結果發現Hispolon確實能對NB4引起細胞毒性、凋亡與週期停滯於G0/G1，且與時間、劑量成相關性。由西方點墨法可得，Hispolon會使細胞凋亡蛋白Bax、caspase 3、8、9表現量增加，與Bcl-2、Bid表現量降低；在細胞週期調控因子方面，會使G0/G1期的cyclin D1、cyclin E、Cdk 2和Cdk 4蛋白表現量減少，p53與p21表現量增加。因此可推測，Hispolon具有抑制人類前骨髓白血病細胞NB4的細胞生長並誘導走向細胞凋亡路徑。 Phellinu merrillii (PM)，is a traditional medicinal plant of oriental people (especially in China, Japan and North Korea ), was demonstrated to exhibit anti-bacterial, antitumour, antifibrotic, antioxidant and anti-inflammatory functions in several studies. Moreover, it has the best anticancer activity in higher fungi as we have already known. Hispolon is a major component of (PM) with great antioxidant activity. And Hispolon had been reported to present apoptotic effect on human epidermoid KB cells, but the exact molecular mechanism of Hispolon on cell death from acute promyelocytic leukemia cell, NB4, is still not clear. Here we investigate it that Hispolon induced cytotoxicity, DNA damage and fragmentation, cell cycle arrest, and apoptosis on NB4 cell. In this study, the dye exclusion (trypan blue) indicated that Hispolon induced cytotoxicity in a time- dependent and dose-dependent manner. Hispolon induced DNA damage and fragmentation was confirmed by DAPI staining and DNA laddering assay. The results demonstrated that Hispolon increased the apoptosis related protein level of caspase 3、8、9 and Bax, and decreased Bcl-2 and Bid. It also promoted the level of cell cycle related proteins such as p53 and p21, and inhibited the expression the levels of cyclin D1、cyclin E、Cdk 2 and Cdk 4. These results suggested that Hispolon inhibited the cell growth, induced apoptosis and might be valuable for therapeutic leukemia disease.
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