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目的:检测培养的婴幼儿血管瘤内皮细胞Fas和Fas Ligand(FasL)的表达量。方法:采用组织块结合酶消化法培养婴幼儿血管瘤内皮细胞,细胞培养成功后,分别运用流式细胞术(flow cytometry,FCM)和荧光定量聚合酶链反应(fluores-cent quantitative polymerase chain reaction,FQ-PCR)2-ΔΔCT法检测培养的血管瘤内皮细胞Fas、FasL和Fas、FasL mRNA表达量,并分别与阳性对照人Jurkat细胞和阴性对照人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)进行比较。结果:FCM检测培养的血管瘤内皮细胞Fas表达率为(90.97±2.36)%,阳性对照Jurkat细胞Fas表达率为(93.87±7.60)%,两者相比无显著差异(P>0.05),但高于阴性对照HUVEC(25.07±7.60)%;FQ-PCR结果显示,血管瘤内皮细胞Fas mRNA的表达量为(1.26±0.72),与阳性对照Jurkat细胞的表达量(1.448±0.059)比值为0.87,差异无显著性(P>0.05),与阴性对照HUVEC的表达量(0.354±0.170)之比为3.56,差异具显著性(P<0.01)。FCM和FQ-PCR检测FasL和FasL mRNA在血管瘤内皮细胞、Jurkat细胞和HUVEC表达量均很低,没有生物学意义。结论:Fas在婴幼儿血管瘤内皮细胞呈高度表达,而FasL的表达量很低,血管瘤内皮细胞凋亡和Fas高表达相关。
Objective: To detect the expression of Fas and Fas Ligand (FasL) in cultured infantile hemangiomas. Methods: Endothelial cells of infantile hemangioma were cultured by tissue block combined with enzymatic digestion. After the cells were cultured successfully, flow cytometry (FCM) and fluorescent-quantitative polymerase chain reaction FQ-PCR) 2-ΔΔCT method was used to detect the expression of Fas, FasL and Fas, FasL mRNA in cultured human hemangioma endothelial cells, and compared with the positive control human Jurkat cells and the negative control human umbilical vein endothelial cells (HUVEC )Compare. Results: The positive rate of Fas expression in endothelial cells was (90.97 ± 2.36)% by FCM, and (93.87 ± 7.60)% in positive control Jurkat cells, but there was no significant difference between the two groups (P> 0.05) (25.07 ± 7.60)%, respectively. The FQ-PCR results showed that the expression of Fas mRNA in endothelial cells was (1.26 ± 0.72), which was significantly higher than that in positive control Jurkat cells (1.448 ± 0.059) , No significant difference (P> 0.05), and the negative control HUVEC expression (0.354 ± 0.170) ratio of 3.56, the difference was significant (P <0.01). FCM and FQ-PCR detection of FasL and FasL mRNA in hemangioma endothelial cells, Jurkat cells and HUVEC expression levels are low, no biological significance. Conclusion: Fas is highly expressed in the endothelial cells of infantile hemangiomas, while the expression of FasL is very low. The expression of Fas is related to the apoptosis of endothelial cells and hemangiomas.