干扰素-γ、干扰素-α2b联合应用诱导MCF-7乳腺癌细胞凋亡及机制

目的 研究干扰素-γ、干扰素-α2b联合应用对MCF-7乳腺癌细胞的诱导凋亡作用及其机制.方法 流式细胞仪（FCM）检测处理后细胞凋亡及Fas/FasL蛋白表达情况.结果 处理后凋亡细胞百分率及FasL的表达增加,随着处理时间的延长,表达有增加趋势,差异有统计学意义（P〈0.05）;Fas的表达无明显变化.结论 干扰素-γ、干扰素-α2b联合应用可诱导MCF-7乳腺癌细胞凋亡,机制可能与上调FasL的表达,加强Fas/FasL之间的交联及使Fas/FasL通路敏感性增加有关.     

Fas/FasL信号传导途径在启动尘肺中的作用机制

目的 探讨Fas/FasL信号传导途径在尘肺发生发展中的作用机制.方法 以中国煤炭工人北戴河疗养院无其他肺部疾病的0 接尘工人及溯、Ⅱ期尘肺患者为研究对象,将支气管肺泡灌洗液(BALF)中的肺泡巨噬细胞(AMs)分5组培养(分别在信号传导途径的不同环节进行抑制),即对照组(未抑制任何途径介导的细胞凋亡过程)、SOD组(在AMs凋亡过程上游保护AMs,抑制AMs细胞膜脂质过氧化过程从而抑制凋亡过程,观察SOD对凋亡细胞的保护作用)、Fas/FasL组(抑制三条传导途径中的另两条即TNFR/TNF-α、TRAILR/TRAIL途径,观察Fas/FasL信号传导途径介导的细胞凋亡情况)、FasL抑制组(同时抑制TNFR/TNF-α、TRAILR/TRAIL和Fas/FasL三条途径,观察Fas/FasL信号传导途径上游抑制后的细胞凋亡情况),caspase-8抑制组(同时抑制TNFR/TNF-α、TRAILR/TRAIL途径和Fas/FasL途径的下游蛋白caspase-8的活性,观察Fas/FasL信号传导途径下游抑制后的细胞凋亡情况).分别采用Western blot(WB)、琼脂糖凝胶电泳(ACE)和TDT介导的dUTP缺口末端标记技术(TUNEL)方法检测肺泡巨噬细胞(AMs)的凋亡情况、DNA片段的改变、信号蛋白Fas、FasL、caspase-8、caspase-3的表达水平.结果 SOD组、FasL抑制组、caspase-8抑制剂组的凋亡指数均低于对照组,差异均有统计学意义.对照组的AMs出现凋亡细胞特征性"梯状带",SOD组未出现,FasL抑制组、caspase-8抑制剂组的DNA条带明显弱于对照组.SOD组的Fas、FasL、cmpase-8、caspase3蛋白的表达均低于对照组,差异均有统计学意义(P＜0.05);FasL抑制组的caspase-8、caspase-3的表达低于Fas/FasL组与对照组,差异均有统计学意义(P＜0.05);caspase-8抑制组的caspase-3表达低于对照组,差异有统计学意义(P＜0.05).结论 尘肺的发生发展可能与Fas/FasL信号传导途径介导的AMs凋亡有关.     

Fas/FasL系统介导的细胞凋亡与卵泡闭锁及稽留流产的关系

Fas/FasL是细胞表面的两种跨膜蛋白。目前研究认为FasL是死亡因子,Fas则是其受体。当一个细胞的FasL与另一个细胞的Fas结合时,可以导致表达Fas的细胞凋亡。Fas又称Apo-1,即CD95分子。人Fas基因定位于10号染色体q23,基因长度约25Kb。人FasL基因定位于1号染色体q23,基因长度约8Kb。Fas/FasL系统与卵泡闭锁生理过程、自身免疫疾病、肿瘤发生、移植物耐受等均有密切关系。近年来细胞凋亡因素导致稽留流产的发生受到人们的关注,而其中Fas/FasL系统在促进绒毛组织细胞凋亡方面占据重要的地位。     

壬基酚对体外培养大鼠睾丸支持细胞凋亡影响的研究

目的探讨壬基酚对大鼠睾丸支持细胞的影响及其作用机制.方法建立体外培养大鼠睾丸支持细胞以及混合培养支持细胞和生精细胞体系,以不同浓度的壬基酚(分别为0.2μg/L、1.0 μg/L、3.0 μg/L、5.0 μg/L)作用于支持细胞和混合培养的支持细胞和生精细胞,采用MTT法测定支持细胞的活性,流式细胞仪测定支持细胞的凋亡率,RT-PCR法测定混合培养的支持细胞和生精细胞中Fas和FasL mRNA表达量的变化.结果随着壬基酚作用剂量的增大和作用时间的延长支持细胞的活性下降,且支持细胞的凋亡率随着壬基酚浓度的增大而提高;混合培养的支持细胞和生精细胞中Fas和FasL mRNA表达量均明显提高.结论壬基酚可引起支持细胞凋亡,进而通过Fas/FasL系统启动生精细胞凋亡.     

亚急性一氧化碳中毒诱导小鼠脑细胞凋亡及相关基因表达

目的 研究亚急性一氧化碳（CO）中毒脑损伤与脑细胞凋亡及其相关基因表达的关系。方法 采用CO100ml/kgip,1次/d。连续7d染毒小鼠，在停止给予CO后第1、3、5、7、14和21天。脑组织在体灌流固定，并用TUNEL法和免疫组织化学法检测大脑皮层和海马凋亡细胞及其相关基因Fas,FasL、Bax蛋白表达情况。结果 亚急性CO中毒小鼠大脑皮层和海马在中毒后第1天可见凋亡细胞，高峰期约在5-7d，持续14d；凋亡相关基因Fas,FasL,Bax蛋白表达在中毒后大量表达，基本与凋亡细胞出现相符。时间稍提前，结论 亚急性CO中毒可诱导神经元细胞凋亡及其相关基因Fas,FasL和Bax蛋白表达，这可能是亚急性CO中毒致迟发性脑损伤的病理机制之一。     

肾癌Fas和FasL蛋白表达异常对其免疫逃避机制的影响

目的探讨肾癌系统表达异常对其免疫逃避机制的影响。方法采用免疫组化方法分析44例肾癌组织及相邻正常肾组织Fas、FasL表达情况;用TUNEL法检测44例肾癌标本中浸润性T淋巴细胞的凋亡。体外培养786-0肾癌细胞株,流式细胞仪(FCM)分析肾癌细胞株Fas、FasL的表达及肿瘤细胞的凋亡;体外培养Fas+的Jurkat细胞,FCM检测其凋亡情况。结果肾癌细胞Fas表达率为22.8%,显著低于其在正常肾组织中表达率(53.8%,P<0.01)。而FasL表达率(46.5%)显著高于其在相邻正常肾组织在的表达(23.2%)。肾癌组织Fas、FasL表达呈负相关(r=-0.46,P<0.05);肾癌浸润性T淋巴细胞凋亡率(33.1%)与肾癌FasL表达呈正相关(r=0.96,P<0.01)。化疗药物5-Fu能上调肾癌细胞株786-0Fas的表达,并增强肾癌细胞对Fas抗体的细胞毒效应,通过抑制肾癌细胞株786-0FasL表达而降低其攻击JurkatT淋巴细胞的能力。讨论肾癌细胞免疫逃避机制包括两方面:一方面通过Fas低表达,能逃避Fas单克隆抗体介导的凋亡;另一方面通过FasL高表达,具有主动攻击免疫淋巴细胞的能力。临床化疗药物5-Fu就Fas系统而言通过上调Fas、抑制FasL的表达发挥一定的疗效。     

超氧化物歧化酶复合酶对尘肺患者肺泡巨噬细胞Fas/FasL信号转导及细胞凋亡的调控作用

目的 探讨超氧化物歧化酶(SOD)复合酶对尘肺患者肺泡巨噬细胞(AM)Fas/FasL信号转导及细胞凋亡的调控作用,为尘肺的早期防治提供依据.方法 收集在某专科医院进行大容量双肺灌洗治疗的0+接尘工人、Ⅰ期、Ⅱ期尘肺患者(共50例)的肺泡灌洗液中的细胞成分,将每例的AM分为非处理组、SOD组、Fas/FasL组.分别取5×106个细胞于DMEM中纯化培养2 h,3组均换新的DMEM,SOD组加SOD复合酶200 U/ml、Fas/FasL组加抗肿瘤坏死因子α((TNF-α)抗体50 ng/ml、抗-TRAIL抗体200 ng/ml后继续培养24 h.取1×106个AM进行细胞裂解,用western blot方法检测Fas、FasL、Caspases-8、Caspases-3蛋白表达,用quantity one 7.0图像分析软件测定各蛋白的相对含量.采用末端脱氧核酸转移酶(TdT)介导的duTP缺口末端标记技术(TFUNEL)检测AM凋亡情况;琼脂糖凝胶电泳法定性检测AM DNA片断化.结果 SOD组、Fas/FasL组凋亡指数分别为(9.50±2.76)%和(14.01±2.56)%,均低于未处理组[(19.18±2.83)%],差异有统计学意义(P＜0.05);未处理组呈现细胞凋亡的特征性DNA梯状带,Fas/FasL组梯状带隐约可见,SOD组未见特征性梯状带.SOD组Ⅰ期、Ⅱ期尘肺患者的Fas、FasL、Caspase-8、Caspase-3蛋白表达水平均低于未处理组和Fas/FasL组,差异有统计学意义(P＜0.05).不同处理组各期尘肺的AM Fas、FasL、Caspase-8、Caspase-3的蛋白表达水平差异均无统计学意义(P＞0.05).结论 SOD复合酶可下调Fas/FasL系统的Fas、FasL、Caspase-8、Caspase-3的蛋白表达水平,抑制AM的凋亡.     

Fas/FasL系统与钙、硒、碘及ROS的关系研究

Fas/FasL系统是介导细胞凋亡的一对膜蛋白,在维持组织发育、控制免疫反应和调节机体生理平衡中起重要作用,并与某些疾病的发生、发展密切相关。近年来研究发现,在细胞凋亡过程中,涉及胞内钙离子的持续上升,不同细胞器内钙离子分布的改变。某些钙离子结合蛋白、钙调素、钙结合蛋白-D 28K等在细胞凋亡过程中可能起着某种调控作用。硒、碘以及活性氧与Fas/FasL途径诱导的细胞凋亡具有密切的联系,可能引起凋亡相关基因表达的改变,激活细胞凋亡程序。     

Fas／FasL参与热诱导大鼠睾丸生精细胞凋亡的调控

目的：探讨睾丸局部加热致大鼠睾丸生精细胞凋亡及Fas/FasL信号通路在生精细胞凋亡调控中的作用。方法16只成年SD雄性大鼠随机均分为加热组和对照组。加热组进行大鼠睾丸43℃水浴15 min;对照组行大鼠睾丸22℃水浴15 min。24 h后进行灌流和内固定,取睾丸。制作5μm的组织切片,采用睾丸原位末端标记法（TUNEL）和免疫组化分析,检测生精细胞凋亡及 Fas 和 FasL 表达。化学发光法检测大鼠血清睾酮水平。结果加热组睾丸生精细胞凋亡率（15.8％±1.6％）较对照组（2.2％±0.5％）显著增加;加热后Fas和FasL在生精细胞和支持细胞表达水平也明显升高;对照组和加热组血清睾酮（T）水平无差异。结论睾丸局部加热诱导生精细胞凋亡,Fas和FasL系统为该凋亡过程的信号通路之一。     

Fas/FasL与宫颈癌关系的研究进展

凋亡是基因控制的、细胞自主有序的死亡,指细胞在一定的生理或病理条件下,遵循自身的程序,自己结束其生命,最后脱落离体裂解为若干凋亡小体而被巨噬细胞吞噬。凋亡在维持机体内环境稳定中起重要作用。Fas与FasL的相互作用是引起细胞凋亡的主要途径之一,深入了解Fas/FasL在宫颈癌中的作用和调控机制,对宫颈癌的诊断、疗效评价及预后判断中具有重要意义。     

Apoptosis occurs in implantation site of the rhesus monkey during early stage of pregnancy.

The exact role of apoptosis that occurs in human placenta during the early stage of pregnancy remains unknown because of the difficulty in obtaining the intact implantation site. In this study, we used rhesus monkey as an animal model to examine apoptosis occurring in the implantation site at various stages of early pregnancy. It was shown that Fas and FasL mRNA and protein were localized in both the chorionic villi and glandular epithelium from day 15 to day 30 of pregnancy. Fas and FasL protein were also expressed in the epithelial plaque on day 15 of pregnancy. In situ 3'-end-labeling results showed that glandular epithelial cells underwent extensive apoptosis with obvious morphological degradation during the early stage of pregnancy. It was found that the cells that were 3'-end-labeled in the chorionic villi and anchoring villi were mainly localized in cytotrophoblast and cytotrophoblast column. It is therefore suggested that in primates apoptosis, which may be involved in the regulation of proliferation of trophoblast villi and degradation of epithelial plaque, as well as remodeling of the glands in the maternal decidua, may play an important role during the early stage of implantation and placentation.     

Fas/Fas L与Survivin在乳腺癌中的研究进展

乳腺癌作为一种激素依赖性实体瘤,它的发生与肿瘤细胞凋亡受阻有关。Fas/Fas L在乳腺癌细胞凋亡中起重要作用,它们在乳腺癌的诊断、治疗及预后方面有很高的临床应用价值。生存素Survivin是凋亡抑制蛋白(inhibitors of apoptosis proteins,IAPs)家族的新成员,参与调控细胞的增殖和凋亡,在人类多种恶性肿瘤中,Survivin高表达并提示预后不良。奉研究对Fas/Fas L和Survivin与乳腺癌的关系研究进展加以综述。     

Fas-independent apoptosis induced by UVC in p53-mutated human epithelial tumor A431 cells through activation of caspase-8 and JNK/SAPK

A431 cells/UVC-induced apoptosis/Caspase 8/Fas/JNK/PAPK. We previously observed that p53-mutated human epithelial tumor A431 cells underwent apoptosis after ultraviolet C (UVC) irradiation through the caspases-8 and -3 pathway. Fas/FasL is known to initiate apoptosis in several cell lines via caspase-8 activation. Then, to determine if Fas/FasL mediates apoptosis in A431. we investigated Fas expression and modulation in UVC-irradiated A431 cells. A431 constitutively expressed Fas, which gradually decreased after UVC-irradiation. Pretreatment with a neutralizing anti-Fas antibody, ZB4, did not abrogate the UVC-induced apoptosis. An agonistic anti-Fas antibody, CH11, very slowly induced apoptosis in A431. suggesting that the constitutively expressed Fas had a low functional potential. Hence, UVC-induced apoptosis in A431 seems to occur independent of the Fas signal. Interestingly, however, a pretreatment with CH11 remarkably potentiated UVC-induced apoptosis. An inhibitor of caspase-8, Ac-IETD-CHO, partially inhibited UVC-induced apoptosis. JNK was phosphorylated immediately after exposure to UVC. prior to apoptotic chromatin condensation. Our data suggest that the activation of caspase-8 occurs independent of Fas upregulation, and that JNK/ SAPK contributes to UVC-induced apoptosis in human epithelial A431 cells.     

硒对实验性自身免疫性甲状腺炎大鼠甲状腺Fas/FasL表达的影响

目的 研究适碘条件下不同硒摄入水平对实验性自身免疫性甲状腺炎(EAT)大鼠甲状腺细胞凋亡蛋白Fas/FasL表达的影响.方法 将32只雌性Lewis大鼠按数字表法随机分成4组,分别设为对照组(C组),模型组(M组),补硒+模型组(Se++M组),低硒+模型组(Se-+M组),在适碘条件下(含10.90 mg/kg)对各组先施加不同水平的硒(Se++M组2 mg/kg,C与M组0.20 mg/kg,Se-+M组0.02 mg/kg)干预2周后,再采用猪甲状腺球蛋白免疫大鼠建立EAT模型,取甲状腺组织制成石蜡切片,采用免疫组织化学检测、比较Fas/FasL表达情况的差异.结果 EAT大鼠甲状腺Fas及FasL较对照均出现表达增加,其中Fas的表达有随补硒水平的升高而下降的趋势,Se++M组吸光度值(0.036±0.004)与M组(0.059±0.006)相比,Fas表达明显受到抑制(q=11.591,P=0.000),与Se-+M组(0.050±0.005)相比,Se++M组Fas呈较低表达水平(q=7.055,P=0.000);但补硒对FasL的表达无明显影响.结论 补硒能减少Fas在甲状腺细胞中的表达,对自身免疫性甲状腺疫病的发展具有抑制作用.     

母胎界面Fas及其配体FasL表达与妊娠期高血压疾病的关系

目的:探讨母胎界面上Fas及其配体FasL表达与妊娠期高血压疾病的关系。方法:采用免疫组化方法对49例产妇母胎界面上Fas及其配体FasL表达进行测定,其中正常孕足月产妇16人,妊娠期高血压疾病轻度子痫前期产妇18名,妊娠期高血压疾病重度子痫前期15名,所有患者均为剖宫产分娩。结果:妊娠期高血压疾病组胎盘滋养细胞Fas表达较正常孕足月组增强并随病情加重更为明显,而FasL着色较正常孕足月组减弱,随病情加重更为明显。在底蜕膜细胞中,随妊娠期高血压疾病的加重FasL表达与正常孕足月相比逐渐减弱。结论:母胎界面Fas及其配体FasL表达紊乱,致使局部免疫反应增强,免役豁免机制被破坏,而高表达的Fas引起胎盘滋养细胞及底蜕膜细胞随之增加,导致胎盘侵蚀异常,血管重铸障碍,可能是妊娠期高血压疾病发病的重要机制之一。     

Regulative effect of IFN-gamma on the Fas/Fas L system of cholangiocarcinoma cells

OBJECTIVE: To detect the regulative effects of IFN-gamma on the expression of Fas and FasL of cholangiocarcinoma cells. METHODS: We studied that the expression of Fas and FasL gene by the human cholangiocarcinoma cell line QBC939 by RT-PCR, Western blot, immunohistochemistry. At the same time, we investigated the regulative effect of IFN-gamma on them. RESULTS: Fas and FasL mRNA and protein were expressed by cholangiocarcinoma cells. We also found IFN-gamma could upregulate the expression of the two genes (P < 0.01). However, IFN-gamma could also downregulate the ability of them to make Jurkat cells apoptotic. With the increasing of dosage and time, the effect was enhanced. CONCLUSIONS: IFN-gamma could regulate the expression of Fas and FasL by cholangiocarcinoma cells, therefore it could reduce the ability of cholangiocarcinoma to occur immune escape. This provides new theoretical basis for immunological therapy of cholangiocarcinoma.     

Fas and FasL promoter polymorphisms and susceptibility to HBV infection: A systematic review and meta-analysis

Apoptosis is a universal cellular defense mechanism against senescent, damaged, genetically mutated, or virally-infected cells. It also is critical for the maintenance of liver health. Fas and FasL system act as a major death pathway that triggers apoptosis cascade in the liver. In this systematic review and meta-analysis, we aimed to investigate the relationship between four major polymorphisms of Fas and FasL genes with susceptibility to or clearance of HBV infection.All the eligible studies were extracted from PubMed and Scopus with no date and language restriction. ORs with 95% CIs were used to evaluate the strength of the association based on the following genetic models: (1) the allelic, (2) the homozygote, (3) the dominant, and (4) the recessive models.Totally 7 related articles were included in this meta-analysis; 5 studies of 7 related articles investigated FasL -844C/T (rs763110) polymorphism, 4 studies investigated FasL IVS2nt-124, 6 studies investigated Fas -670 A/G (rs1800682), and 4 studies investigated Fas -1377 A/G (rs2234767) polymorphism. This meta-analysis showed that there is no statistically significant association between the risk or clearance of HBV infection and four studied Fas and FasL polymorphisms in their allelic comparison or genetic models.Fas -670, Fas -1377, FasL -124, and FasL -844 polymorphisms did not show any significant association with the clearance or risk of HBV infection. Therefore, it seems that susceptibility to HBV infection or clearance of it is not affected by Fas and FasL genetic polymorphisms. But, to reach a definitive conclusion, further studies with a larger sample size of different ethnicity are still needed.