Jiang Yu-Ping, Wu Xiao-Hua, Xing Han-Ying, DU Xing-Yan
Department of Obstetrics and Gynecology, Second Hospital of Hebei Medical University, Shijiazhuang 050000, China.
Chin Med J (Engl). 2007 Jul 20;120(14):1251-5.
In a previous study, we have verified that CXCR4 expression is correlated with tumor aggressive progression and poor prognosis in patients with epithelial ovarian cancer. The aim of this study was to explore the effect of CXCL12-CXCR4 axis on the metastasis of human ovarian cancer.
The expressions of CXCR4 and CXCL12 mRNA and protein in human ovarian cancer cell line CAOV-3 was detected by RT-PCR and immunocytochemistry. Methythiazolyltetrazolium (MTT) was used to analyze the effect of different concentrations of CXCL12 on the proliferation of CAOV-3 cells. Transwell invasion chamber and matrigel were used to evaluate the effect of various concentrations of CXCL12 and ascites on the migration and invasion of CAOV-3 cells. The expressions of integrin beta(1) and vascular endothelial growth factor-C (VEGF-C) mRNA were detected by RT-PCR. Data were analyzed using ANOVA by SAS 6.12.
Under serum-free suboptimal culture conditions, CXCL12 (100 ng/ml) significantly enhanced the proliferation of CAOV-3 cells compared with the control and 10 ng/ml CXCL12 groups (0.428 +/- 0.051 vs. 0.325 +/- 0.045 and 0.328 +/- 0.039, P < 0.05). This enhancing effect of CXCL12 was significantly inhibited by 10 microg/ml neutralizing CXCR4 antibody or 1 microg/ml CXCR4 antagonist AMD3100. However, 10 microg/ml neutralizing CXCR4 antibody could not inhibit cell proliferation without CXCL12. The levels of migration and invasion of the CAOV-3 cells treated with 100 ng/ml CXCL12 were significantly higher than those in the control (migration: 523.3 +/- 25.2 vs 108.0 +/- 7.2; invasion: 39.3 +/- 4.0 vs. 4.0 +/- 1.0). The enhancing effect of CXCL12 on cell migration and invasion increased with the concentration of CXCL12 (100 ng/ml vs10 ng/ml: migration, 523.3 +/- 25.2 vs 211.7 +/- 24.7; invasion, 39.3 +/- 4.0 vs 15.7 +/- 3.1, P < 0.05), and was strongly inhibited by 10 microg/ml neutralizing CXCR4 antibody or 1 microg/ml AMD3100. The number of migrated and invading cells in the CAOV-3 added with ascites was significantly higher than those in the 100 ng/ml CXCL12 group (migration: 706.6 +/- 30.6 vs 523.3 +/- 25.2, invasion: 61.7 +/- 7.6 vs 39.3 +/- 4.0, P < 0.05). The level of integrin beta(1) mRNA was greatly increased at 3 hours after being treated with CXCL12 (0.53 +/- 0.10 vs. 1.53 +/- 0.16, P < 0.05), and VEGF-C mRNA displayed significant augment at 24 hours after being treated with CXCL12 (0.52 +/- 0.09 vs 1.11 +/- 0.15, P < 0.05).
CXCL12 and its receptor CXCR4 can promote the proliferation, migration, invasion of ovarian cancer cell line CAOV-3 and enhance its secretion of integrin beta(1) and VEGF-C. These effects can be inhibited by neutralizing CXCR4 antibody or AMD3100. CXCL12-CXCR4 axis plays an important role in ovarian cancer growth and metastasis.
在之前的一项研究中，我们已经证实CXCR4表达与上皮性卵巢癌患者的肿瘤侵袭性进展和不良预后相关。本研究的目的是探讨CXCL12 - CXCR4轴对人卵巢癌转移的影响。
采用逆转录 - 聚合酶链反应（RT - PCR）和免疫细胞化学法检测人卵巢癌细胞系CAOV - 3中CXCR4和CXCL12 mRNA及蛋白的表达。用甲基噻唑基四氮唑（MTT）分析不同浓度CXCL12对CAOV - 3细胞增殖的影响。使用Transwell侵袭小室和基质胶评估不同浓度CXCL12和腹水对CAOV - 3细胞迁移和侵袭的影响。通过RT - PCR检测整合素β1和血管内皮生长因子 - C（VEGF - C）mRNA的表达。数据采用SAS 6.12软件进行方差分析。
在无血清次优培养条件下，与对照组和10 ng/ml CXCL12组相比，CXCL12（100 ng/ml）显著增强了CAOV - 3细胞的增殖（0.428±0.051对0.325±0.045和0.328±0.039，P < 0.05）。10 μg/ml中和性CXCR4抗体或1 μg/ml CXCR4拮抗剂AMD3100可显著抑制CXCL12的这种增强作用。然而，10 μg/ml中和性CXCR4抗体在无CXCL12时不能抑制细胞增殖。用100 ng/ml CXCL12处理的CAOV - 3细胞的迁移和侵袭水平显著高于对照组（迁移：523.3±25.2对108.0±7.2；侵袭：39.3±4.0对4.0±1.0）。CXCL12对细胞迁移和侵袭的增强作用随CXCL12浓度的增加而增加（100 ng/ml对10 ng/ml：迁移，523.3±25.2对211.7±24.7；侵袭，39.3±4.0对15.7±3.1，P < 0.05），并被10 μg/ml中和性CXCR4抗体或1 μg/ml AMD3100强烈抑制。添加腹水的CAOV - 3细胞中迁移和侵袭细胞的数量显著高于100 ng/ml CXCL12组（迁移：706.6±30.6对523.3±25.2，侵袭：61.7±7.6对39.3±4.0，P < 0.05）。用CXCL12处理3小时后，整合素β1 mRNA水平大幅升高（0.53±0.10对1.53±0.16，P < 0.05），用CXCL12处理24小时后，VEGF - C mRNA显著增加（0.52±0.09对1.11±0.15，P < 0.05）。
CXCL12及其受体CXCR4可促进卵巢癌细胞系CAOV - 3的增殖、迁移、侵袭，并增强其整合素β1和VEGF - C的分泌。这些作用可被中和性CXCR4抗体或AMD3100抑制。CXCL12 - CXCR4轴在卵巢癌的生长和转移中起重要作用。
