The induction of orphan nuclear receptor nur77 expression by n-butylenephthalide as pharmaceuticals on hepatocellular carcinoma cell therapy
The induction of orphan nuclear receptor nur77 expression by n-butylenephthalide as pharmaceuticals on hepatocellular carcinoma cell therapy
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DOI：
10.1124/mol.107.044800
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发表时间：
2008-10-01
影响因子：
3.6
通讯作者：
Harn, Horng-Jyh
中科院分区：
文献类型：
--
作者：
Chen, Yi-Lin;Jian, Min-Hui;Harn, Horng-Jyh
从当归氯仿提取物中分离得到的正丁基苯酞(BP)，已被用于检测其对多形性胶质母细胞瘤脑瘤的抗肿瘤作用，但对其对肝细胞癌细胞的抗肿瘤作用知之甚少。分别用N-丁叉对苯二酚和赋形剂处理肝癌细胞株HepG2和J5，检测细胞存活率和细胞凋亡率。研究孤儿受体家族Nurr1、Nor-1和Nur77等与细胞凋亡相关的基因和蛋白的表达，以及N-丁叉苯酞在体内异种移植模型中的作用。在25µg/ml的浓度下，N-丁基苯酞对这两种细胞的生长均有抑制作用。此外，N-丁基苯酞诱导的细胞凋亡可能与Nur77从胞核移位到胞浆，导致细胞色素c释放和caspase-3依赖的细胞凋亡有关。与N-丁叉苯酞相关的肿瘤细胞凋亡与磷脂酰肌醇3-激酶/蛋白激酶B(AKT)/糖原合成酶-3β相关，而不是丝裂原激活的蛋白激酶或蛋白激酶C途径。阻断AKT激活可增强对细胞增殖的抑制和对磷酸化的Bcl2和Nur77蛋白的诱导。此外，BP通过野生型cAMP反应元件结合蛋白(CREB)进入肿瘤细胞而增加的细胞凋亡率被CREB的显性磷酸化位点突变所抑制。这一发现提示CREB途径也参与了BP诱导的肿瘤细胞凋亡。在HepG2和J5移植瘤中，N-丁叉对苯二酚有相似的抗肿瘤作用。N-丁叉对苯二酚在体内外均可诱导肝癌细胞发生凋亡，提示该化合物有可能用于改善肝癌细胞的预后。
N-Butylidenephthalide (BP), isolated from the chloroform extract of Angelica sinensis, has been examined for its antitumor effects on glioblastoma multiforme brain tumors; however, little is known about its antitumor effects on hepatocellular carcinoma cells. Two hepatocellular carcinoma cell lines, HepG2 and J5, were treated with either N-butylidenephthalide or a vehicle, and cell viability and apoptosis were evaluated. Apoptosis-related mRNA and proteins expressed, including orphan receptor family Nurr1, NOR-1, and Nur77, were evaluated as well as the effect of N-butylidenephthalide in an in vivo xenograft model. N-Butylidenephthalide caused growth inhibition of both the cell lines at 25 mu g/ml. Furthermore, N-butylidenephthalide- induced apoptosis seems to be related to Nur77 translocation from nucleus to cytosol, which leads to cytochrome c release and caspase-3-dependent apoptosis. N-Butylidenephthalide-related tumor apoptosis was associated with phosphatidylinositol 3-kinase/protein kinase B (AKT)/glycogen synthase kinase-3 beta rather than the mitogen-activated protein kinase or protein kinase C pathway. Blockade of AKT activation enhanced proliferation inhibition and the induction of phosphor-Bcl-2 and Nur77 proteins. Besides, the increasing apoptosis by BP via transfection wild-type cAMP-response element-binding protein (CREB) into tumor cell was suppressed by dominant phosphorylation site mutation of CREB. This finding suggested CREB pathway was also partly involved in tumor apoptosis caused by BP. Administration of N-butylidenephthalide showed similar antitumoral effects in both HepG2 and J5 xenograft tumors. N-Butylidenephthalide induced apoptosis in hepatocellular carcinoma cells, both in vitro and in vivo, suggesting a potential clinical use of this compound for improving the prognosis of hepatocellular carcinoma cells.