Scala Stefania, Giuliano Paola, Ascierto Paolo A, Ieranò Caterina, Franco Renato, Napolitano Maria, Ottaiano Alessandro, Lombardi Maria L, Luongo Monica, Simeone Ester, Castiglia Daniele, Mauro Francesca, De Michele Ileana, Calemma Rosa, Botti Gerardo, Caracò Corrado, Nicoletti Gianfranco, Satriano Rocco A, Castello Giuseppe
Department of Clinical Immunology, National Cancer Institute, G. Pascale Foundation, Naples, Italy.
Clin Cancer Res. 2006 Apr 15;12(8):2427-33. doi: 10.1158/1078-0432.CCR-05-1940.
The chemokine receptor CXCR4 was identified as an independent predictor of poor prognosis in primary melanoma. The aim of the study was to investigate the role of CXCR4 in human melanoma metastases.
CXCR4 expression was evaluated in melanoma metastases and in metastatic cell lines through immunohistochemistry, immunoblotting, immunofluorescence, and reverse transcription-PCR. The function of CXCR4 was tested in the presence of the ligand, CXCL12, through induction of extracellular signal-regulated kinase-1 and -2 (Erk-1 and -2) phosphorylation, proliferation, apoptosis, and migration capabilities.
CXCR4 expression was detected in 33 out of 63 (52.4%) metastases from cutaneous melanomas. Metastatic melanoma cell lines expressed cell surface CXCR4; PES 43, Alo 40, and COPA cell lines showed the highest levels of CXCR4 (>90% of positive cells); PES 41, Alo 39, PES 47, POAG, and CIMA cell lines showed low to moderate degrees of expression (5-65% of positive cells). Other chemokine receptors, CCR7 and CCR10, were detected on the melanoma cell lines; CXCL12 activated Erk-1 and Erk-2, the whose induction was specifically inhibited by AMD3100 treatment. CXCL12 increased the growth in PES 41, PES 43, and PES 47 cells under suboptimal (1% serum) and serum-free culture conditions; AMD3100 (1 mumol/L) inhibited the spontaneous and CXCL12-induced proliferation. No rescue from apoptosis was shown but PES 41, PES 43, and PES 47 cells migrate toward CXCL12.
These findings indicate that CXCR4 is expressed and active in human melanoma metastases, suggesting that active inhibitors such as AMD3100 may be experienced in human melanoma.
趋化因子受体CXCR4被确定为原发性黑色素瘤预后不良的独立预测指标。本研究旨在探讨CXCR4在人类黑色素瘤转移中的作用。
通过免疫组织化学、免疫印迹、免疫荧光和逆转录聚合酶链反应，评估CXCR4在黑色素瘤转移灶和转移细胞系中的表达。在存在配体CXCL12的情况下，通过诱导细胞外信号调节激酶1和2（Erk-1和Erk-2）磷酸化、增殖、凋亡和迁移能力来测试CXCR4的功能。
在63例皮肤黑色素瘤转移灶中的33例（52.4%）检测到CXCR4表达。转移性黑色素瘤细胞系表达细胞表面CXCR4；PES 43、Alo 40和COPA细胞系显示CXCR4水平最高（>90%阳性细胞）；PES 41、Alo 39、PES 47、POAG和CIMA细胞系显示低至中度表达（5-65%阳性细胞）。在黑色素瘤细胞系中检测到其他趋化因子受体CCR7和CCR10；CXCL12激活Erk-1和Erk-2，AMD3100处理可特异性抑制其诱导。在次优（1%血清）和无血清培养条件下，CXCL12增加了PES 41、PES 43和PES 47细胞的生长；AMD3100（1μmol/L）抑制自发和CXCL12诱导的增殖。未显示凋亡的挽救，但PES 41、PES 43和PES 47细胞向CXCL12迁移。
这些发现表明CXCR4在人类黑色素瘤转移中表达并具有活性，提示活性抑制剂如AMD3100可能用于人类黑色素瘤治疗。
