摘要
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 examine the expression of Fas and Fas ligand（FasL） in cultured infantile hemangioma endothelial cells （IHECs）. Methods： IHECs were isolated and cultured by explant combined with trypsin digested techniques. Then the expression of Fas and FasL on cultured endothelial cells （ECs） were detected by flow eytometry （FCM） and fluorescent quantitative polymerase chain reaction （FQ-PCR） 2^-△△CT method,and the results in ECs were compared with those in Jurkat cells （positive control） and Human umbilical vein endothelial cells（HUVECs, negative control）. Results： The positive rate of Fas in cultured IHECs was （90. 97±2.36）% as detected by FCM,similar to that in the Jurkat cells （[93.87±1.64]% ,P〉0.05） and significantly higher than that in the HUVECs （P〈0.01）. Fas mRNA expression in the cultured IHECs was 1. 260±0. 721 as detected by FQ-PCR,and was 1. 448±0. 059 in Jurkat cells,with their ratio being 0.87 and with on significant difference found between the 2 group（P〉0.05） ; the ratio of IHECs to HUVECs （0. 354±0. 170） was 3.56 and there was significant difference between the 2 groups （P〈0.01）. The expression of FasL mRNA in cultured IHECs,Jurkat cells and HUVECs was too low as detected by FCM and FQ-RCR to have any biological significance. Conclusion： Fas mRNA is highly expressed in cultured IHECs and FasL is rarely expressed in IHECs, which indicates that apoptosis of IHECs is associated with high expression of Fas in IHECs.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2008年第11期1296-1299,共4页
Academic Journal of Second Military Medical University
基金
国家自然科学基金(30571928)~~
