Targeting the ATF-1/CREB transcription factors by single chain Fv fragment in human melanoma: potential modality for cancer therapy.
Targeting the ATF-1/CREB transcription factors by single chain Fv fragment in human melanoma: potential modality for cancer therapy.
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DOI：
10.1615/critrevimmunol.v21.i1-3.180
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发表时间：
2001
影响因子：
1.3
通讯作者：
D. Jean;M. Bar‐eli
中科院分区：
文献类型：
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作者：
D. Jean;M. Bar‐eli
转录激活因子-1（ATF-1）和cAMP反应元件（CRE）结合蛋白（CREB）参与cAMP和Ca ~（2+）诱导的转录激活。转录因子ATF-1和CREB的表达在转移性黑素瘤细胞中上调。然而，ATF-1/CREB的过表达如何促进转移表型的获得尚不清楚。以前，我们证明了通过显性阴性形式的CREB（KCREB）淬灭转移性黑色素瘤细胞中的CREB活性，导致其在裸鼠中的致瘤性和转移潜力降低。我们确定了两种机制来解释CREB/ATF-1的过度表达如何有助于转移表型。第一种是CREB/ATF-1通过调节CREB依赖的金属蛋白酶MMP-2和粘附分子MCAM/MUC 18基因的表达在侵袭中发挥重要作用。在第二种机制中，CREB和ATF-1作为人黑色素瘤细胞的存活因子。在此，使用抑制性抗ATF-1单链抗体片段（ScFv）的细胞内表达来研究破坏ATF-1活性的效果。MeWo黑色素瘤细胞中ScFv抗ATF-1的细胞内表达导致CRE依赖性启动子激活显着减少。此外，在黑色素瘤细胞中表达抗ATF-1单链抗体抑制其在裸鼠中的致瘤性和转移潜力。抗ATF-1单链抗体使黑色素瘤细胞在体外对毒胡萝卜素诱导的凋亡敏感，并在皮下移植到裸鼠的肿瘤中引起大量凋亡，证实AFT-1/CREB作为人黑色素瘤细胞的存活因子。这些研究证明了抗ATF-1单链抗体在体内作为肿瘤生长和转移的抑制剂的潜在用途。
Activating transcription factor-1 (ATF-1) and cAMP-responsive element (CRE)-binding protein (CREB) have been implicated in cAMP and Ca2+-induced transcriptional activation. The expression of the transcription factors ATF-1 and CREB is upregulated in metastatic melanoma cells. However, how overexpression of ATF-1/CREB contributes to the acquisition of the metastatic phenotype is unclear. Previously we demonstrated that quenching of CREB activity in metastatic melanoma cells by means of a dominant-negative form of CREB (KCREB) led to a decrease in their tumorigenicity and metastatic potential in nude mice. We identified two mechanisms that explain how overexpression of CREB/ATF-1 contributes to the metastatic phenotype. The first is one in which CREB/ATF-1 play an essential role in invasion by regulating the CRE-dependent expression of the metalloproteinase MMP-2 and the adhesion molecule MCAM/MUC18 genes. In the second mechanism, CREB and ATF-1 act as survival factors for human melanoma cells. Here, the effect of disrupting ATF-1 activity was investigated using intracellular expression of an inhibitory anti-ATF-1 single chain antibody fragment (ScFv). Intracellular expression of ScFv anti-ATF-1 in MeWo melanoma cells caused significant reduction in CRE-dependent promoter activation. In addition, expression of ScFv anti-ATF-1 in melanoma cells suppressed their tumorigenicity and metastatic potential in nude mice. ScFv anti-ATF-1 rendered the melanoma cells susceptible to thapsigargin-induced apoptosis in vitro and caused massive apoptosis in tumors transplanted subcutaneously into nude mice, confirming that AFT-1/CREB act as survival factors for human melanoma cells. These studies demonstrate the potential usage of ScFv anti-ATF-1 as an inhibitor of tumor growth and metastasis of solid tumors in vivo.