Zhang Ran, Banik Naren L, Ray Swapan K
Department of Neurosciences, Medical University of South Carolina, Charleston, SC 29425, USA.
Neurochem Res. 2007 Dec;32(12):2194-202. doi: 10.1007/s11064-007-9417-7. Epub 2007 Jul 7.
Phosphatase and tension homolog located on chromosome ten (PTEN) is a tumor suppressor as it negatively regulates activation of Akt. Mutation or deletion of PTEN has been found in as high as 80% of glioblastomas, which harbor aberrant cell signaling passing through the phosphatidylinositol-3-kinase (PI3K) and Akt (PI3K/Akt) survival pathway. Glioblastoma cells without functional PTEN are not easily amenable to apoptosis. We investigated the possibility of modulation of signal transduction pathways for induction of apoptosis in human glioblastoma T98G (PTEN-harboring) and U87MG (PTEN-deficient) cell lines after treatment with the combination of all-trans retinoic acid (ATRA) and interferon-gamma (IFN-gamma). Treatment with ATRA plus IFN-gamma stimulated PTEN expression and suppressed Akt activation in T98G cells, whereas no PTEN expression but Akt activation in U87MG cells under the same conditions. Pretreatment of U87MG cells with the PI3K inhibitor LY294002 could prevent Akt activation. Interestingly, ATRA plus IFN-gamma could significantly decrease cell viability and increase morphological features of apoptosis in both cell lines. Combination of ATRA and IFN-gamma showed more efficacy than IFN-gamma alone in causing apoptosis that occurred due to increases in Bax:Bcl-2 ratio, mitochondrial release of cytochrome c, and caspase-3 activity. Luciferase reporter gene assay showed that combination of ATRA and IFN-gamma significantly down regulated transcriptional activity of the nuclear factor kappa B (NF-kappaB), a survival signaling factor, in U87MG cells. Thus, combination of ATRA and IFN-gamma caused significant amounts of apoptosis in T98G cells due to suppression of the PI3K/Akt survival pathway while the same treatment caused apoptosis in U87MG cells due to down regulation of the NF-kappaB activity. Therefore, the combination of ATRA and IFN-gamma could modulate different survival signal transduction pathways for induction of apoptosis and should be considered as an effective therapeutic strategy for controlling the growth of both PTEN-harboring and PTEN-deficient glioblastomas.
位于10号染色体上的磷酸酶和张力同源物(PTEN)是一种肿瘤抑制因子,因为它能负向调节Akt的激活。在高达80%的胶质母细胞瘤中发现了PTEN的突变或缺失,这些肿瘤存在通过磷脂酰肌醇-3-激酶(PI3K)和Akt(PI3K/Akt)生存通路的异常细胞信号传导。缺乏功能性PTEN的胶质母细胞瘤细胞不易发生凋亡。我们研究了在全反式维甲酸(ATRA)和干扰素-γ(IFN-γ)联合处理后,调节信号转导通路以诱导人胶质母细胞瘤T98G(携带PTEN)和U87MG(PTEN缺陷)细胞系凋亡的可能性。ATRA加IFN-γ处理刺激了T98G细胞中PTEN的表达并抑制了Akt的激活,而在相同条件下U87MG细胞中没有PTEN表达但有Akt激活。用PI3K抑制剂LY294002预处理U87MG细胞可以阻止Akt激活。有趣的是,ATRA加IFN-γ能显著降低两种细胞系的细胞活力并增加凋亡的形态学特征。ATRA和IFN-γ联合使用在诱导凋亡方面比单独使用IFN-γ更有效,这是由于Bax:Bcl-2比值增加、细胞色素c从线粒体释放以及caspase-3活性增加所致。荧光素酶报告基因检测表明,ATRA和IFN-γ联合使用能显著下调U87MG细胞中生存信号因子核因子κB(NF-κB)的转录活性。因此,ATRA和IFN-γ联合使用通过抑制PI3K/Akt生存通路在T98G细胞中导致大量凋亡,而相同处理通过下调NF-κB活性在U87MG细胞中导致凋亡。因此,ATRA和IFN-γ联合使用可以调节不同的生存信号转导通路以诱导凋亡,应被视为控制携带PTEN和PTEN缺陷的胶质母细胞瘤生长的有效治疗策略。