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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 |
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(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) |
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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。 |
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). |
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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 |