S1P/S1PR对人卵巢癌SKOV3细胞的促血管生成作用的研究

目的：探究鞘氨醇-1-磷酸/鞘氨醇-1-磷酸受体（S1P/S1PR）对人卵巢癌SKOV3细胞促血管生成作用的影响和机制。方法：管腔形成实验探究S1P对人卵巢癌SKOV3细胞的促血管生成作用的影响；实时定量聚合酶链式反应（qRT-PCR）检测S1P处理后的人卵巢癌SKOV3细胞中血管生成因子白细胞介素8（IL-8）、IL-6、血管内皮生长因子（VEGF）的变化情况；设计合成小干扰RNA（siRNA）干扰序列基因沉默SKOV3细胞中S1PR1、S1PR2和S1PR3的表达，蛋白质印迹（Western-blot）和qRT-PCR检测SKOV3细胞中S1PR的下调结果；qRT-PCR检测S1PR对SKOV3细胞IL-8、IL-6、VEGF表达的影响。结果：管腔形成实验结果显示，S1P处理后的SKOV3细胞培养上清液重悬人脐静脉内皮细胞融合细胞（EA.hy926），其管腔形成的数量多于对照组（t=3.667，P=0.021），表明S1P促进了人卵巢癌细胞SKOV3的促血管形成能力。S1P处理后的SKOV3细胞中血管生成因子IL-8、IL-6、VEGF mRNA的表达水平较对照组升高，差异有统计学意义（均P＜0.05）。S1PR1和S1PR3基因沉默可显著降低SKOV3细胞中IL-8、IL-6、VEGF的mRNA表达水平，差异有统计学意义（均P＜0.05），而S1PR2基因沉默后IL-8、IL-6、VEGF的mRNA变化不明显。结论：S1P通过S1PR1/3上调人卵巢癌SKOV3细胞中IL-8、IL-6、VEGF的表达，从而增强了SKOV3细胞的促血管生成作用，S1P/S1PR通路有望成为抑制卵巢癌生长的治疗新靶点。

Study on Effect of S1P/S1PR on Angiogenesis in SKOV3 Ovarian Cancer Cells

Objective：The study was to explore the effect and mechanism of S1P/S1PR on angiogenesis in human ovarian cancer cell line (SKOV3). Methods：Tube formation assay was to explore the angiogenetic effect of S1P on ovarian cancer cells. QRT-PCR was used to identify IL-8, IL-6 and VEGF mRNA expression changes in SKOV3 ovarian cancer cells incubation with S1P. SKOV3 cells were transfected with siRNA interference sequences silencing S1PR1, S1PR2 and S1PR3 gene. S1PR mRNA levels were determined by qRT-PCR and protein levels were determined by western-blot. IL-8, IL-6 and VEGF change levels in S1PR gene silencing ovarian cancer cell lines were detected by qRT-PCR. Results：The tube formation essay of human umbilical vein endothelial cells was significantly increased by the culture supernatants of ovarian cancer cells treated with S1P (t=-3.667, P=0.021). The results showed that S1P promoted the pro-angiogenic ability of SKOV3. The mRNA expression of IL-8, IL-6 and VEGF were significantly increased by S1P in SKOV3 cells (P＜0.05). The mRNA expression levels of IL-8, IL-6 and VEGF in SKOV3 cells were significantly decreased in S1PR1 and S1PR3 gene silencing cells (all P＜0.05) and were not significantly changed in S1PR2 gene silencing cells. Conclusions：The study indicated that S1P could affect ovarian cancer cells through S1PR1/3 to promote ovarian cancer angiogenesis and IL-8, IL-6, VEGF may play a role in this process. S1P/S1PR pathway is expected to become a new target for ovarian cancer therapy.