Fas、FasL在婴幼儿血管瘤内皮细胞的表达
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国家自然科学基金(30571928).


Expression of Fas and Fas ligand in endothelial cells of infantile hemangioma
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Supported by National Natural Science Foundation of China(30571928).

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    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 (fluorescent 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。

    Abstract:

    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 cytometry (FCM) and fluorescent quantitative polymerase chain reaction (FQ-PCR) 2-ΔΔCT 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.

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  • 收稿日期:2008-04-30
  • 最后修改日期:2008-09-15
  • 录用日期:2008-09-17
  • 在线发布日期: 2008-11-19
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