Ianaro Angela, Tersigni Mariaroberta, Belardo Giuseppe, Di Martino Silvana, Napolitano Maria, Palmieri Giuseppe, Sini MariaCristina, De Maio Anna, Ombra MariaNeve, Gentilcore Giuseppina, Capone Mariaelena, Ascierto MariaLibera, Satriano Rocco Alfredo, Farina Benedetta, Faraone-Mennella MariaRosaria, Ascierto Paolo Antonio, Ialenti Armando
Department of Experimental Pharmacology, University of Naples Federico II, Via D. Montesano 49, 80131 Naples, Italy.
Cancer Lett. 2009 Feb 18;274(2):331-6. doi: 10.1016/j.canlet.2008.09.038. Epub 2008 Nov 11.
Melanoma is the most aggressive form of skin cancer, it originates from melanocytes and its incidence has increased in the last decade. Recent advances in the understanding of the underlying biology of the progression of melanoma have identified key signalling pathways that are important in promoting melanoma tumourigenesis, thus providing dynamic targets for therapy. One such important target identified in melanoma tumour progression is the Nuclear Factor-kappaB (NF-kappaB) pathway. In vitro studies have shown that NF-kappaB binding is constitutively elevated in human melanoma cultures compared to normal melanocytes. It has been found that a short cell-permeable peptide spanning the IKK-beta NBD, named NBD peptide, disrupted the association of NEMO with IKKs in vitro and blocked TNFalpha-induced NF-kappaB activation in vivo. In the present study we investigated the effect of the NBD peptide on NF-kappaB activity and survival of A375 human melanoma cells. We found that NBD peptide is able to inhibit the proliferation of A375 cells, which present constitutively elevated NF-kappaB levels. Inhibition of cell proliferation by NBD peptide was associated with direct inhibition of constitutive NF-kappaB DNA-binding activity and induction of apoptosis by activation of caspase-3 as confirmed by the cleavage and consequently inactivation of poly (ADP ribose) polymerase (PARP-1) known as the best marker of this process.
黑色素瘤是最具侵袭性的皮肤癌形式，它起源于黑素细胞，且在过去十年中其发病率有所上升。对黑色素瘤进展的潜在生物学机制理解的最新进展，已确定了在促进黑色素瘤肿瘤发生中起重要作用的关键信号通路，从而为治疗提供了动态靶点。在黑色素瘤肿瘤进展中确定的一个此类重要靶点是核因子-κB（NF-κB）通路。体外研究表明，与正常黑素细胞相比，人黑色素瘤培养物中NF-κB的结合持续升高。已发现一种跨越IKK-β NBD的短细胞穿透肽，名为NBD肽，在体外破坏了NEMO与IKK的结合，并在体内阻断了肿瘤坏死因子α（TNFα）诱导的NF-κB激活。在本研究中，我们研究了NBD肽对A375人黑色素瘤细胞的NF-κB活性和存活的影响。我们发现NBD肽能够抑制A375细胞的增殖，这些细胞呈现出持续升高的NF-κB水平。NBD肽对细胞增殖的抑制与对组成型NF-κB DNA结合活性的直接抑制以及通过激活半胱天冬酶-3诱导细胞凋亡有关，这一点通过聚（ADP核糖）聚合酶（PARP-1）的切割及随后的失活得到证实，PARP-1是这一过程的最佳标志物。
