Cong Rihong, Sun Qingmin, Yang Li, Gu Haijuan, Zeng Ying, Wang Bin
Department of Pharmacology, Nanjing Medical University, Nanjing, PR China.
J Exp Clin Cancer Res. 2009 Sep 7;28(1):124. doi: 10.1186/1756-9966-28-124.
Vasculogenic mimicry (VM) was increasingly recognized as a form of aggressive melanoma acquiring blood supply. Genistein had attracted much attention as a potential anticancer agent. Therefore, we examined the effect of Genistein on VM in human uveal melanoma cells.
VM structure was detected by periodic acid-Schiff (PAS) staining for uveal melanoma C918 cells cultured on the three-dimensional type I collagen gels after exposed to Genistein. We used reverse transcription polymerase chain reaction (RT-PCR) and Western Blot analysis to examine the effect of Genistein on vascular endothelial cadherin (VE-cadherin) mRNA and protein expression. The nude mice models of human uveal melanoma C918 cells were established to assess the number of VM using immunohistochemical and PAS double-staining.
Genistein inhibited the survival of C918 cells in vitro. The ectopic model study showed that VM in tumor tissue sections were significantly reduced by Genistein in vivo. In vitro, the VM structure was found in control, 25 and 50 microM Genistein-treatment groups but not in 100 and 200 microM. RT-PCR and Western Blot showed that 100 and 200 microM concentration of Genistein could significantly decrease VE-cadherin mRNA and protein expression of C918 cells compared with control (P < 0.05). However, the 25 and 50 microM Genistein slightly decreased the VE-cadherin level in vitro (P > 0.05).
Genistein inhibits VM formation of uveal melanoma cells in vivo and in vitro. One possible underlying molecular mechanism by which Genistein could inhibit VM formation of uveal melanoma is related to down-regulation of VE-cadherin.
血管生成拟态（VM）作为侵袭性黑色素瘤获取血供的一种形式，越来越受到认可。染料木黄酮作为一种潜在的抗癌药物已引起广泛关注。因此，我们研究了染料木黄酮对人葡萄膜黑色素瘤细胞中VM的影响。
对在三维I型胶原凝胶上培养的葡萄膜黑色素瘤C918细胞暴露于染料木黄酮后，采用过碘酸希夫（PAS）染色检测VM结构。我们使用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹分析来检测染料木黄酮对血管内皮钙黏蛋白（VE-钙黏蛋白）mRNA和蛋白表达的影响。建立人葡萄膜黑色素瘤C918细胞的裸鼠模型，采用免疫组织化学和PAS双重染色评估VM的数量。
染料木黄酮在体外抑制C918细胞的存活。异位模型研究表明，染料木黄酮在体内可显著减少肿瘤组织切片中的VM。在体外，对照组、25和50微摩尔/升染料木黄酮处理组中发现有VM结构，但100和200微摩尔/升处理组中未发现。RT-PCR和蛋白质免疫印迹显示，与对照组相比，100和200微摩尔/升浓度的染料木黄酮可显著降低C918细胞的VE-钙黏蛋白mRNA和蛋白表达（P<0.05）。然而，25和50微摩尔/升染料木黄酮在体外轻微降低了VE-钙黏蛋白水平（P>0.05）。
染料木黄酮在体内和体外均抑制葡萄膜黑色素瘤细胞的VM形成。染料木黄酮抑制葡萄膜黑色素瘤VM形成的一种可能潜在分子机制与VE-钙黏蛋白的下调有关。
