Characterization of the lipolytic pathways that mediate free fatty acid release during Fas/CD95-induced apoptosis
Marı́a IturraldeJulián PardoEsther LacasaGloria del BarrioM.A. ÁlavaAndrés PiñeiroJavier NavalAlberto Anel
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bcl-2 암유전자는 여러 가지 자극에 의한 apoptosis를 차단함으로써 유전자 이상을 가진 세포가 계속 생존하면서 유전자 변이가 누적되는 결과를 초래한다고 알려져 있다. 한편 c-myc 암유전자는 세포증식과 apoptosis를 유도하는 이중적 기능을 가지고 있으며 생존 신호가 결여될 경우에는 오히려 세포의 apoptosis를 유발한다고 알려져 있다. 그러나 c-myc과 bcl-2가 동시에 발현되면 bcl-2는 c-myc의 세포증식 작용은 영향을 주지 않고 apoptosis만을 선택적으로 차단함으로써 유전자 변이 세포의 생존 뿐만 아니라 증식을 촉진하는 것으로 관찰되었다. 자궁경부암에서 c-myc과 bcl-2 발현에 관한 개별적 보고는 있었으나 이들 두 유전자의 동시발현 및 이들 유전자들이 실제 암조직상에서 세포증식 및 apoptosis에 어떠한 영향을 미치는가에 관한 연구는 보고된 것이 없다. 따라서 본 연구에서는 자궁경부암 발생 과정중에서 bcl-2 및 c-myc 발현과 세포증식, apoptosis와의 상관관계를 알아보고자 하였다. 본 연구에서는 10개의 정상 자궁경부조직, 30개의 자궁경부 상피내종양 조직, 20개의 자궁경부암조직에서 bcl-2와 c-myc에 대한 면역조직화학 검사를 시행하였으며 세포증식과 apoptosis는 각각 Ki-67 면역조직화학적 방법과 TUNEL 방법으로 확인하였다. 또한 환자의 임상병리학적 인자들과의 상관관계도 알아보았다. 정상 자궁경부, 자궁경부 상피내종양, 자궁경부암조직 중 자궁경부암조직에서만 bcl-2와 c-myc 단백이 각각 35%와 50%에서 관찰되었으며, 또한 bcl-2와 c-myc의 동시발현이 25%에서 관찰되었다. 세포증식 지수(상피세포 100개중 Ki-67양성 세포수)는 정상 자궁경부, 상피내종양, 자궁경부암으로 진행되면서 10.2, 24.1, 59.7, 71.2로 유의하게 증가하는 양상을 보였으며(p<0.01), apoptosis 지수(상피세포 100개중 apoptosis 세포수)도 0, 0.33, 1.85, 3.89로 점차 증가하는 양상을 보였다(p<0.01). 또한 세포증식 지수와 apoptosis 지수와는 높은 상관관계(r=0.7451, p=0.0002)를 나타내었다. 그러나 자궁경부암 조직중 bcl-2 발현군과 비발현군간에 apoptosis지수에는 차이가 없었으며(p=0.4765), c-myc 발현군과 비발현군간에도 세포증식 지수에는 차이가 없었다(p=0.6891). 또한 bcl-2와 c-myc의 동시 발현군과 나머지 군간에도 증식지수와 apoptosis 지수에 차이가 없었다(각각 p=0.6311 및 p=0.7600). 한편 bcl-2와 c-myc의 동시 발현과 잘 알려져 있는 자궁경부암의 임상병리학적 예후인자들(종양 크기, FIGO 임상병기, 림프절 전이등)과는 유의한 상관관계가 없었다. 이상과 같은 결과에서 자궁경부암발생 과정에서 세포증식과 apoptosis는 병변의 등급과 비례하여 증가하고 apoptosis는 세포증식과 관련된 변화로 사료되었다. 한편 bcl-2와 c-myc과 발현은 자궁경부암에서만 관찰되는 유전자 변이로서 자궁경부 상피내종양의 발생과 진행과정에는 영향을 미치지 않으며, 또한 자궁경부암 조직에서도 암조직 전체의 세포증식 및 apoptosis와는 관련이 없을 것으로 사료되었다.
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瞄准：学习是否消炎痛(IND )(NS ) ，一个非选择的 cyclooxygenase (艇长) 禁止者或 NS-398，一个 COX-2-selective 禁止者，在人的结肠癌房间导致 apoptosis 并且 apoptosis 相关的基因和小径它被包含。方法：人的结肠癌 Caco-2 房间与也被对待：安慰剂， IND (0.05-0.5 mmol/L ) 或为 1， 5 和 18 h 的 NS (0.01-0.2 mmol/L ) 。我们然后学习了：(1 ) 由 TUNEL 方法的细胞死亡，(2 ) 用 DNA 微数组的 96 apoptosis 相关的基因的 mRNA 表示，(3 ) 选择 apoptosis 的表示由西方的弄污联系了蛋白质。结果：IND 和 NS 以一种剂量依赖者方式在 Caco-2 房间的 30%-50% 导致了 apoptosis。IND (为 1 h 的 0.1 mmol/L ) 在四个家庭的显著地起来调整的 pro-apoptotic 基因：(1 ) TNF 受体和 ligand，(2 ) Caspase，(3 ) Bcl-2 并且(4 ) Caspase 招募领域。NS 治疗起来调整的类似的 pro-apoptotic 基因作为 IND。另外，国际机场家庭的 IND 也下面调整的 anti-apoptotic 基因。结论：(1 ) 非选择并且在以一种剂量依赖者方式的结肠癌房间的 COX-2-selective NSAID induce apoptosis。(2 ) 两 NSAID 由激活二条主要 apoptotic 小径导致 apoptosis：死亡受体小径(包含的 TNF-R ) 和 mitochondrial 小径。(3 ) IND 由起来调整的 pro-apoptotic 基因和下面调整的 anti-apoptotic 基因导致 apoptosis，当时 NS 仅仅起来调整 pro-apoptotic 基因。(4 ) 在由 NSAID 的结肠癌房间的 apoptosis 的正式就职可以部分地解释，他们结肠癌生长上的禁止的行动。
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There are two distinct modes of cell death: necrosis and apoptosis. Apoptosis is an active process that may require gene expression and protein synthesis, and for this reason apoptosis has been called programmed cell death. Cancer cells in culture exposed to several anticancer drugs show morphological and biochemical characteristics consistent with apoptosis. This observation has possible impact on chemotherapeutic treatment of cancer. In this article we survey apoptosis, with emphasis on the relationship between apoptosis and anticancer drugs.
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AIM To investigate the effect of cisplatin on apoptosis in Scaber cell. METHODS The apoptotic cells were detected by TUNEL,HE,eletronic micrpscopy. RESULTS Treatment of Scaber cells with CDDP resulted in characteristics typical of apoptosis. CDDP induced apoptosis of Scaber cells in time and concentration dependent manner. To further investigate the mechanism of apoptosis induced by CDDP, the expressions and activity of apoptosis associated proteins such as bcl 2, bax and caspase 3 were examined using S P method.The results showed: CDDP caused time and concentration dependent decreases in bcl 2 and increased in bax proteins.CDDP bcl 2 and its translocation to perinuclei and nuclei. The expression of caspase 3 in Scaber cell were determined during apoptosis induced by CDDP. CONCLUSION Our investigetion showed that the apoptosis induced by CDDP is related to the increase of bax protein, and the decrease of bcl 2 protein. and its translocation to perinuclei and nuclei.
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瞄准：为了在人的胰腺的癌症房间调查 apoptosis，导致了由 Triptolide (TL ) ，和在 caspase-3' bcl-2 和 bax 的这 apoptosis 和表情之间的关系。方法：人的胰腺的癌症房间线 SW1990 在这研究的 DMEM 媒介是有教养的。MTT 试金被用来决定房间生长禁止的率在试管内。流动血细胞计数和 TUNEL 试金被用来在 TL 治疗前后检测人的胰腺的癌症房间的 apoptosis。RT-PCR 被用来检测联系 apoptosis 的基因 caspase-3' bcl-2 和 bax 的表示。结果：TL 在剂量禁止了人的胰腺的癌症房间的生长 -- 并且时间依赖者举止。TL 导致了人的胰腺的癌症房间经历 apoptosis 与典型地 apoptotic 特征。TUNEL 试金为 12 h 和 24 h 与 40 ng/mL TL 在人的胰腺的癌症房间的治疗以后显示出那，人的胰腺的癌症房间的 apoptotic 率显著地增加了。RT-PCR 表明了那 caspase-3， bax 在当 bcl-2 mRNA 不是时，与 TL 对待的 SW1990 房间是显著地起来调整的。结论：TL 能在人的胰腺的癌症房间导致 apoptosis。这 apoptosis 可以被调停由起来调整联系 apoptosis 的 caspase-3 和 bax 基因的表示。
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To explore the role of Fas gene in rhG-CSF-induced apoptosis.Mediated by lipofection (DOTAP), Fas cDNA was transferred into HL-60 cells. In situ hybridization, Western blotting and FCM analysis were used to demonstrate the successful transfection. The apoptosis percentages of HL-60 and Fas transfected HL-60 cells were compared after coculturing with rhG-CSF at the same final concentration.After cocultured with rhG-CSF (10 ng/ml) for 96 hours, the apoptosis percentage of Fas transferred HL-60 cells was significantly higher than that of HL-60 cells, and the Fas expressions could be up-regulated by rhG-CSF.Fas is an apoptosis-induction oncogene, and rhG-CSF induces apoptosis of HL-60 cells through Fas signalling pathway.
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Autoimmune lymphoproliferative syndrome
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瞄准：为了调查低紧张超声(美国) ，在人的胃的癌房间和它的潜在的机制并且到导致了 apoptosis 建议一条新治疗学的途径到胃的癌。方法：人的 SGC-7901 胃的癌房间是有教养的在试管内并且在照耀以后与不同孵化时间在不同紧张为 10 min 由低紧张美国照耀。词法变化在显微镜下面被检验，尝试平底锅蓝色染色然后早 apoptotic 房间的百分比被流动血细胞计数(FCM ) 与两倍染色荧光黄异硫氰酸盐(FITC ) 检测 -Annexin V/propidium 碘化物(PI ) 。二维的电气泳动(2DE ) 被用来让蛋白质侧面和一些蛋白质在美国照耀被帮助矩阵的激光解吸附作用 / 电离 time-of-flight 团分光术(MALDI-TOF-MS ) 识别以后，不同地表示了。功能的分析被执行调查导致美国的房间 apoptosis 的机制。结果：apoptotic 房间的百分比在美国照耀以后增加了大约 10%(12.0 W/cm2， 12 h 文化) 。在照耀美国的房间的早 apoptosis 和第二等的坏死的百分比与增加的美国紧张增加了。而且， apoptotic 房间在美国照耀以后与增加的文化时间增加了并且在大约 12 h 到达了它的最大值。几新蛋白质在美国照耀以后出现了并且起来或下面调整了超过 2 次。一些热吃惊蛋白质(HSP ) 被发现与模仿房间的 apoptosis 的信号过程被联系。结论：低紧张美国能在人的胃的癌房间导致 apoptosis。导致美国的 apoptosis 与美国紧张 / 文化时间有关。导致美国的 apoptosis 可能是 caspases 依赖的并且 endoplasmic (嗯) 被触发压力的 apoptosis 可以也贡献它。Proteomic 试验性的系统在在美国照耀以后在癌房间发现蛋白质改变是有用的，帮助开发新癌症治疗。
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The aim of the investigation was to estimate the relation of CD95 and DR3 receptors activation with apoptosis level of naive cytotoxic Т-lymphocytes (nCТL) in children with acute infectious mononucleosis (AIM). Materials and Methods. The test materials were peripheral blood samples of healthy children and children with AIM. nCTL were isolated by negative immunоmagnetic separation. Specific activation of CD95 and DR3 receptors was performed using monoclonal antibodies. An apoptosis level and expression of receptors were studied by flow cytometry. Results. The percentage of cell apoptosis decreased in children with AIM in freshly isolated nCTL, as well as in CD95 receptor activation compared to healthy children. nCTL apoptosis in healthy children regardless of culture conditions was accompanied by the reduced quantity of CD95+DR3– cells and CD95 expression density on their surface. In children with AIM the decrease of these indices required CD95 activation. Compared to healthy children, the percentage of CD95+DR3+ cells in children with AIM decreased in CD95 activation. In CD95 receptor activation in healthy children and children with AIM, the content of CD95+DR3+ cells correlated directly with an apoptosis level. DR3 receptor activation was accompanied neither by nCTL apoptosis level change nor the changed content of DR3+ cells in both healthy children and children with AIM. Conclusion. nCTL are less sensitive to apoptosis in children with AIM compared to healthy children. DR3 receptor activation results in no change of nCTL apoptosis level both in healthy children and children with AIM. CD95 activation in patients with AIM is accompanied by increased resistance of CD95+DR3– cells to apoptosis and the susceptibility to apoptosis of CD95+DR3+ cells. The evaluation of nCTL susceptibility to CD95-induced apoptosis in AIM can serve as a subtest to assess the state of a cell component of immune system.
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