摘要
AIM： To investigate whether alphastatin could inhibit human gastric cancer growth and furthermore whether sphingosine kinase （SPK） activity is involved in this process. METHODS： Using migration assay, MTT assay and Matrigel assay, the effect of alphastatin on vascular endothelial cells （ECs） was evaluated in vitro. SPK and endothelial differentiation gene （EDG）-1, -3, -5 mRNAs were detected by reverse transcription-polymerase chain reaction （RT-PCR）. SPK activity assay was used to evaluate the effect of alphastatin on ECs. Matrigel plug assay in nude mice was used to investigate the effect of alphastatin on angiogenesis in vivo. Female nude mice were subcutaneously implanted with human gastric cancer cells （BGC823） for the tumor xenografts studies. Micro vessel density was analyzed in Factor Ⅷ-stained tumor sections by the immunohistochemical SP method. RESULTS： In vitro, alphastatin inhibited the migration and tube formation of ECs, but had no effect on proliferation of ECs. RT-PCR analysis demonstrated that ECs expressed SPK and EDG-1, -3, -5 mRNAs. In vivo, alphastatin sufficiently suppressed neovascularization of the tumor in the nude mice. Daily administration of alphastatin produced significant tumor growth suppression. Immunohistochemical studies of tumor tissues revealed decreased micro vessel density in alphastatin-treated animals as compared with controls. CONCLUSION： Downregulating ECs SPK activity may be one of the mechanisms that alphastatin inhibits gastric cancer angiogenesis. Alphastatin might be a useful and relatively nontoxic adjuvant therapy in the treatment of gastric cancer.
瞄准：调查 alphastatin 是否能禁止人的胃的癌症生长并且而且鞘氨醇激酶(SPK ) 活动是否涉及这个过程。方法：用迁居试金， MTT 试金和 Matrigel 试金，脉管的 endothelial (EC ) 上的 alphastatin 的效果是评估在试管内。SPK 和内皮区别基因(EDG )-1,-3,-5 mRNAs 被反向的抄写聚合酶链反应(RT-PCR ) 检测。SPK 活动试金被用来在 EC 上评估 alphastatin 的效果。在裸体老鼠的 Matrigel 塞子试金被用来在血管生成在活体内上调查 alphastatin 的效果。雌裸体老鼠皮下地为肿瘤异种皮移植研究与人的胃的癌症房间(BGC823 ) 被植入。微容器密度被免疫在因素染色 VIII 的肿瘤节分析组织化学的 SP 方法。结果：在试管内， alphastatin 禁止了迁居和 EC 的试管形成，但是没在 EC 的增长上有效果。RT-PCR 分析证明 EC 表示了 SPK 和 EDG-1， -3,-5 mRNAs。在活体内， alphastatin 足够地在裸体老鼠压制了肿瘤的 neovascularization。alphastatin 的每日的管理生产了重要肿瘤生长抑制。肿瘤纸巾的 Immunohistochemical 研究作为与控制相比在对待 alphastatin 的动物揭示了减少的微容器密度。结论：Downregulating EC SPK 活动可以是 alphastatin 禁止的机制之一胃的癌症血管生成。Alphastatin 可能是在胃的癌症的治疗的有用、相对无毒的辅助治疗。
