Overexpression of both CXC chemokine receptor 4 and vascular endothelial growth factor proteins predicts early distant relapse in stage II-III colorectal cancer patients
Overexpression of both CXC chemokine receptor 4 and vascular endothelial growth factor proteins predicts early distant relapse in stage II-III colorectal cancer patients
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DOI：
10.1158/1078-0432.ccr-05-2142
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发表时间：
2006-05-01
影响因子：
11.5
通讯作者：
Scala, S
中科院分区：
文献类型：
--
作者：
Ottaiano, A;Franco, R;Scala, S
目的：CXC趋化因子受体4(CXCR4)和血管内皮生长因子(VEGF)参与恶性肿瘤的转移过程。然而，目前还没有关于这些分子在结直肠癌中的生物学关系的数据。我们研究了CXCR4和VEGF的表达是否可以预测复发，并在体外评价了CXCR4在促进结直肠癌细胞克隆生长、VEGF分泌和细胞间黏附分子-1(ICAM-1)表达方面的作用。实验设计：用免疫组织化学方法研究CXCR4和VEGF在结直肠癌组织和LOVO、HT29和SW620结直肠癌细胞系中的作用。采用卡方检验分析患者和肿瘤患者基线特征的相关性。用酶联免疫吸附试验检测CXCL12诱导的血管内皮生长因子的分泌。流式细胞仪检测CXCL12对ICAM-1表达的影响。用克隆形成实验检测CXCL12诱导的集落生长。体外给予CXCR4的非竞争性拮抗剂AMD3100可阻止CXCL12引起的功能效应。结果：研究对象为2003年1月至2004年1月的72例患者。CXCR4在16例(22.2%)肿瘤中不表达，在31例(43.0%)肿瘤中有50%的细胞表达。在17例(23.6%)肿瘤中未见表达，16例(22.2%)肿瘤中有50%的细胞表达，39例(54.2%)肿瘤中有50%的细胞表达。CXCR4的表达与淋巴转移、淋巴结转移密切相关(P=0.0393)。血管内皮生长因子的表达与肿瘤侵袭(P=0.0386)和淋巴结转移(P=0.0044)显著相关。单因素分析显示，美国癌症分期联合委员会(P=0.0016)、血管内皮生长因子表达(P=0.0450)、CXCR4表达(P=0.0428)和血管内皮生长因子/CXCR4表达(P=0.0004)对无瘤生存期有显著的预后价值。多因素分析证实了美国癌症分期联合委员会的预测能力以及伴随的和高表达的血管内皮生长因子和CXCR4。对于原发肿瘤在50%的细胞中表达CXCR4和VEGF的患者的预后尤其不利(复发患者的中位无病生存期为5.8个月；复发风险比为8.23；95%可信区间为7.24-14.29)。在克隆形成实验中，CXCL12(20 ng/mL/d)在第7天和第14天显著增加了SW620、HT29和LOVO细胞的克隆数。同样，CXCL12能够刺激SW620、HT29和LOVO细胞分泌血管内皮生长因子，并上调ICAM-1。结论：CXCR4和血管内皮生长因子的同时高表达是结直肠癌早期远处复发的独立预测因子。CXCR4触发了一系列现象，包括刺激克隆生长、诱导血管内皮生长因子释放和ICAM-1上调。这些数据支持抑制CXCR4以阻止结直肠癌转移的发展。
Purpose: CXC chemokine receptor 4 (CXCR4) and vascular endothelial growth factor (VEGF) are implicated in the metastatic process of malignant tumors. However, no data are currently available on the biological relationship between these molecules in colorectal cancer. We studied whether CXCR4 and VEGF expression could predict relapse and evaluated in vitro the contribution of CXCR4 in promoting clonogenic growth,VEGF secretion, and intercellular adhesion molecule-1 (ICAM-1) expression of colorectal cancer cells.Experimental Design: CXCR4 and VEGF were studied in colorectal cancer tissues and in Lovo, HT29, and SW620 colorectal cancer cell lines by immunohistochemistry. Correlations with baseline characteristics of patients and tumors were analyzed by chi(2) test. VEGF secretion induced by CXCL12 was measured by ELISA. The effect of CXCL12 on ICAM-1 expression was evaluated by flow cytometry. Clonogenic growth induced by CXCL12 was determined by clonogenic assays. Functional effects induced by CXCL12 were prevented by the administration in vitro of AMD3100, a bicyclam noncompetitive antagonist of CXCR4.Results: Seventy-two patients, seen between January 2003 and January 2004, were studied. CXCR4 was absent in 16 tumors (22.2%); it was expressed in 50% of cells in 31 (43.0%) tumors. VEGF was absent in 17 (23.6%) tumors; it was expressed in: 50% of cells in 16 (22.2%) tumors and in >50% of cells in 39 (54.2%) tumors. There was a significant association between CXCR4 expression and lymph, nodal status (P = 0.0393). There were significant associations between VEGF and tumor invasion (P = 0.0386) and lymph nodal involvement (P = 0.0044). American Joint Committee on Cancer stage (P = 0.0016),VEGF expression (P = 0.0450), CXCR4 expression (P = 0.0428), and VEGF/CXCR4 expression (P = 0.0004) had a significant prognostic value for disease-free survival with univariate analysis. The predictive ability of the American Joint Committee on Cancer stage and of the concomitant and high expression of VEGF and CXCR4 was confirmed by multivariate analysis. Prognosis is particularly unfavorable for patients whose primary tumors express CXCR4 and VEGF in >50% of cells (median disease-free survival in relapsed patients, 5.8 months; hazard ratio of relapse, 8.23; 95% confidence interval, 7.24-14.29). In clonogenic assays, CXCL12 (20 ng/mL/d) significantly increased the number of clones in SW620, HT29, and Lovo cells at 7 and 14 days. Again, CXCL12 was able to stimulate VEGF secretion in SW620, HT29, and Lovo cells as well as up-regulated ICAM-1. These effects were prevented by the administration of AMD3100 (1 mu mol/L).Conclusions: We have shown that concomitant and high expression of CXCR4 and VEGF is a strong and independent predictor of early distant relapse in colorectal cancer. CXCR4 triggers a plethora of phenomena, including stimulation of clonogenic growth, induction of VEGF release, and ICAM-1 up-regulation. These data support the inhibition of CXCR4 to prevent the development of colorectal cancer metastasis.