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Fas、FasL在婴幼儿血管瘤内皮细胞的表达
廖洪跃1,邢新1*,欧阳天祥2,郭伶俐1,李军辉1,薛春雨1,王晓云1,袁斯明3
0
(1.第二军医大学长海医院整形外科,上海 200433;2.上海交通大学新华医院整形外科,上海 200092;3.南京军区总医院烧伤整形科,南京 210002)
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。
关键词:  血管瘤  内皮细胞  Fas  FasL
DOI:10.3724/SP.J.1008.2008.01296
投稿时间:2008-04-30修订日期:2008-09-15
基金项目:国家自然科学基金(30571928).
Expression of Fas and Fas ligand in endothelial cells of infantile hemangioma
LIAO Hong-yue 1,XING Xin 1* ,OUYANG Tian-xiang 2,GUO Ling-li 1 ,LI Jun-hui 1 ,XUE Chun-yu 1,WANG Xiao-yun 1 ,YUAN Si-ming3
(1.Department of Plastic Surgery,Changhai Hospital,Second Military Medical University,Shanghai 200433,China ;2.Department of Plastic Surgery,Xinhua Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 2000923.;Department of Burn and Plastic Surgery,General Hospital,PLA Nanjing Military Area Command,Nanjing 210002)
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.
Key words:  hemangioma  endothelial cells  Fas  FasL