黃烷酮類化合物導致骨肉瘤細胞死亡的探討及其2’-羥基黃烷酮引起骨肉瘤細胞株143 B產生細胞凋亡的機制與探討

林孟賢

doi:10.6834/CSMU.2011.00018

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黄烷酮类化合物导致骨肉瘤细胞死亡的探讨及其2’-羟基黄烷酮引起骨肉瘤细胞株143 B产生细胞凋亡的机制与探讨

The Study of Flavanones-induced Death in Osteosarcoma Cells and the Mechanism of 2’-hydroxyflavanone-induced Apoptosis in Osteosarcoma 143 B Cell

林孟贤 , 硕士　　导师：谢易修;陈霈霓　　

繁体中文 DOI： 10.6834/CSMU.2011.00018

骨肉瘤；细胞凋亡； osteosarcoma ； apoptosis

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摘要 │ 参考文献 (43) │ 国际计量

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Flavonoids are a group of polyphenol compounds that exist extensively in dietary foods of vegetables and fruits, and the biological effects of flavonoids are anti-oxidant, anti-microbial, and anti-inflammatory activities. Recently, more and more studies have showed that flavonoids have the anti-tumor and anti-carcinogenesis effects. Therefore, the anti-tumor effects of flavanone （flavanone、2'-OH flavanone、3'-OH flavanone、4'-OH flavanone、5'-OH flavanone、6'-OH flavanone、7'-OH flavanone） in the osteosarcoma cell line （U-2 OS、143 B、MG-63、SaOS2） were analyzed. The result of MTT assay indicated that effect of flavanone、2'-OH flavanone and 3'-OH flavanone showed the most potent cytotoxic effect in the osteosarcoma cell line, and we also found that 2'-OH flavanoneare was the most potent cytotoxic compound. The result of cell’s live and death number calculation showed that 2'-OH flavanone induced the death number increasing of osteosarcoma cells. The further way, we found the phenomenon of chromatin condensation、mitochondria membrane potential change and diploid cell （sub-G1 phase） appearance after treatment to 143 B cell. Then, we analyzed the expression of apoptotic protein molecule by western blotting. The result showed the activation of caspase-3、caspase-8 and caspase-9. Therefore, the activation of caspase evidenced the occurrence of apoptosis. The result showed that 2'-OH flavanone increased the expression of TRAIL（TNF-released apoptosis-inducing ligand） and DR5 （Death receptor 5）. On the other hand, the result also showed that 2'-OH flavanone induced cytochrome c was released from mitochondria. We also found the anti-apoptotic proteins （Bcl-2、Bcl-xL）were inhibited, and the pro-apoptotic proteins（Bid、Bax、Bcl-xS）were activated. Therefore, the 2'-OH flavanone induced the 143 B cell apoptosis through TRAIL-DR5 pathway.Finally, we conducted the in vivo experiment（143 B-BALB/c nude mice）. The result showed that tumor growth were suppressed by 2'-OH flavanone. These results indicated that 2'-OH flavanone activated the extrinsic pathway and the intrinsic pathway through TRAIL-DR5 pathway, and it had potent effect to tumor growth in in vivo model. Therefore, 2'-hydroxyflavanone is a potent biological activity compound for the development of cancer chemotherapy.

参考文献 ( 43 ) 〈TOP〉

Wiley SR, Schooley K, Smolak PJ, Din WS, Huang CP, Nicholl JK, Sutherland GR, Smith TD, Rauch C, Smith CA, et al.（1995）. Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity. 3(6):673-82.
连结：
Pitti RM, Marsters SA, Ruppert S, Donahue CJ, Moore A, Ashkenazi A.（1996）. Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family. J Biol Chem. 271(22):12687-90.
连结：
Mariani SM, Matiba B, Armandola EA, Krammer PH.（1997）. Interleukin 1 beta-converting enzyme related proteases/caspases are involved in TRAIL-induced apoptosis of myeloma and leukemia cells. J Cell Biol. 137(1):221-9.
连结：
Liu X, Kim CN, Yang J, Jemmerson R, Wang X. （1996）. Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell. 86(1):147-57.
连结：
Chauhan D, Pandey P, Ogata A, Teoh G, Krett N, Halgren R, Rosen S, Kufe D, Kharbanda S, Anderson K. （1997）. Cytochrome c-dependent and -independent induction of apoptosis in multiple myeloma cells. J Biol Chem. 272(48):29995-7.
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DOI可强化引用精确性、增强学术圈连结，并给予使用者跨平台的良好使用经验，目前在全世界已有超过五千万个对象申请DOI。 如想对DOI的使用与概念有进一步了解，请参考 （ ） 。

DOI可强化引用精确性、增强学术圈连结，并给予使用者跨平台的良好使用经验，目前在全世界已有超过五千万个对象申请DOI。如想对DOI的使用与概念有进一步了解，请参考（）。