Nuclear Factor κB-Mediated Induction of Flice-Like Inhibitory Protein Prevents Tumor Necrosis Factor α-Induced Apoptosis in Rat Granulosa Cells1
Nuclear Factor κB-Mediated Induction of Flice-Like Inhibitory Protein Prevents Tumor Necrosis Factor α-Induced Apoptosis in Rat Granulosa Cells1
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核因子 κB 介导的 Flice 样抑制蛋白诱导可防止肿瘤坏死因子 α 诱导的大鼠颗粒细胞凋亡1
DOI：
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
B. Tsang
中科院分区：
文献类型：
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作者：
Chao Wu Xiao;E. Asselin;B. Tsang
摘要本研究旨在探讨肿瘤坏死因子α (TNFα)对大鼠颗粒细胞中抗凋亡类fliclike inhibitory protein （FLIP）的作用及其调控作用。在无血清RPMI中培养未成熟大鼠颗粒细胞，不含或存在TNFα (20 ng/ml)，环己亚胺（CHX, 10 μg/ml）， SN50（核因子κB [NFκB]易位特异性抑制剂，100或200 μg/ml），或这些的组合。（以SN50突变的失活肽SM50作为对照。）用Western blot和电泳迁移量转移法分别测定抑制剂κB （IκB总形式和磷酸化形式）和NFκB结合能力。采用原位TUNEL法检测细胞凋亡，采用半定量逆转录-聚合酶链反应法检测FLIP mRNA水平。单独TNFα不能诱导颗粒细胞死亡，但在环己亚胺的作用下，凋亡细胞数量明显增加。TNFα显著上调短链FLIP （FLIPS）的表达，而不上调长链FLIP （FLIPL）的表达。TNFα诱导i - κ b磷酸化和活化。SN50可减弱tnf α诱导的FLIPS表达，增强tnf α诱导的细胞凋亡。通过FLIPS反义表达下调tnf α诱导的FLIPS可增强tnf α诱导的细胞凋亡。克隆并测序了与小鼠具有高度同源性（85%）的大鼠FLIPS全长。这些发现表明，除了其促凋亡功能外，TNFα还可以诱导维持卵泡发育的细胞内存活因子。tnf α诱导、nf κ b介导的FLIPS表达是颗粒细胞命运的决定因素。
Abstract The purpose of the present studies was to examine the role and regulation of the antiapoptotic Flice-like inhibitory protein (FLIP) in rat granulosa cells by tumor necrosis factor α (TNFα) in vitro. Granulosa cells from immature rats primed with eCG were cultured in serum-free RPMI in the absence or presence of TNFα (20 ng/ml), cycloheximide (CHX, 10 μg/ml), SN50 (a specific inhibitor of nuclear factor κB [NFκB] translocation, 100 or 200 μg/ml), or a combination of these. (SM50, a mutated inactive peptide of SN50, was used as control.) Inhibitor κB (IκB; total and phosphorylated forms) and NFκB binding abilities were measured by Western blot and electrophoretic mobility shift assay, respectively. Apoptosis was assessed by in situ TUNEL assay, whereas FLIP mRNA levels were determined by semiquantitative reverse transcriptase-polymerase chain reaction. TNFα alone failed to induce granulosa cell death but significantly increased the apoptotic cell number in the presence of cycloheximide. TNFα significantly up-regulated the expression of the short form of FLIP (FLIPS) but not the long form (FLIPL). TNFα induced IκB phosphorylation and NFκB activation. SN50, but not SM50, attenuated TNFα-induced FLIPS expression and enhanced TNFα-induced apoptosis. Down-regulation of TNFα-induced FLIPS by FLIPS antisense expression enhanced TNFα-induced apoptosis. A full length of rat FLIPS, with high homology to mouse FLIPS (85%), had been cloned and sequenced. These findings suggest that, in addition to its proapoptotic function, TNFα can induce an intracellular survival factor for the maintenance of follicular development. TNFα-induced, NFκB-mediated FLIPS expression is a determinant of granulosa cell fate.
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DOI：
10.1073/pnas.94.21.11333
发表时间：
1997-10-14
影响因子：
11.1
作者：
Han, DKM;Chaudhary, PM;Hood, L
通讯作者：
Hood, L
影响因子：
4.8
作者：
HUGHES, FM;GOROSPE, WC
通讯作者：
GOROSPE, WC
影响因子：
4.8
作者：
Antti Kaipia;S. Chun;K. Eisenhauer;Aaron J. W. Hsueh
通讯作者：
Antti Kaipia;S. Chun;K. Eisenhauer;Aaron J. W. Hsueh
DOI：
10.1073/pnas.86.7.2336
发表时间：
1989-04-01
影响因子：
11.1
作者：
OSBORN, L;KUNKEL, S;NABEL, GJ
通讯作者：
NABEL, GJ
肿瘤坏死因子-α 在卵泡发育、排卵和黄体寿命中的潜在作用。
DOI：
10.1016/s0739-7240(96)00094-x
发表时间：
1997
期刊：
Domestic animal endocrinology.
影响因子：
--
作者：
Terranova,PF
通讯作者：
Terranova,PF