Lebovic Dan I, Baldocchi Russell A, Mueller Michael D, Taylor Robert N
Reproductive Endocrinology Division, Department of Obstetrics and Gynecology, University of Michigan, Ann Arbor, Michigan, USA.
Fertil Steril. 2002 Oct;78(4):849-54. doi: 10.1016/s0015-0282(02)03319-8.
Interleukin (IL)-1beta, a product of activated peritoneal macrophages, is a central cytokine coordinating neovascularization and monocyte chemotaxis in endometriotic implants. To evaluate the effects of this cytokine on normal endometrial stromal cells and endometriotic stromal cells we performed cDNA expression array analyses before and after exposure to IL-1beta.
Nested case-control study of women with and without laparoscopic evidence of endometriosis.
Reproductive endocrinology clinic at a university hospital.
PATIENT(S): Endometriosis and normal endometrial biopsies from eight patients were used to prepare stromal cell cultures from which mRNA was extracted.
INTERVENTION(S): None.
MAIN OUTCOME MEASURE(S): Commercially available expression arrays (Atlas Human cDNA Expression Array, Clontech, representing 597 individual genes) were used to screen for mRNAs whose expression was affected by 12 hours of exposure to IL-1beta (10 ng/mL). Northern blotting and subsequent quantitative densitometric evaluation was done to confirm steady-state levels of Tob-1 mRNA transcripts.
RESULT(S): Array analyses revealed a cell-cycle regulatory gene, Tob-1, which was differentially expressed by the two cell types after incubation with IL-1beta. Tob-1 was reduced 48% in endometriotic stromal cells exposed to IL-1beta, but there was only a 16% reduction in normal endometrial stromal cells. Replicate Northern analyses (n = 4) showed that exposure to IL-1beta for 12 hours resulted in a 25% +/- 5% diminution of Tob-1 mRNA in endometriotic stromal cells. In contrast, no significant decrease (<3%) was observed in IL-1beta exposed normal endometrial stromal cells.
CONCLUSION(S): Tob-1, a cell-cycle inhibitor gene is differentially responsive to IL-1beta in endometriotic stromal cells compared to normal endometrial stromal cells. IL-1beta down-regulated Tob-1 in endometriotic stromal cells, but had no significant effect on normal endometrial stromal cells. Our results suggest that IL-1beta promotes growth of endometriotic lesions through inhibition of Tob-1. These findings are the first to associate IL-1beta with an alteration of cell-cycle gene expression in cells derived from endometriotic implants.
白细胞介素（IL）-1β是活化腹膜巨噬细胞的产物，是协调子宫内膜异位症种植体中新血管形成和单核细胞趋化性的核心细胞因子。为了评估这种细胞因子对正常子宫内膜基质细胞和子宫内膜异位症基质细胞的影响，我们在暴露于IL-1β之前和之后进行了cDNA表达阵列分析。
对有和没有腹腔镜检查证据的子宫内膜异位症女性进行巢式病例对照研究。
大学医院的生殖内分泌诊所。
使用8名患者的子宫内膜异位症和正常子宫内膜活检组织制备基质细胞培养物，并从中提取mRNA。
无。
使用市售表达阵列（Atlas Human cDNA Expression Array，Clontech，代表597个个体基因）筛选其表达受暴露于IL-1β（10 ng/mL）12小时影响的mRNA。进行Northern印迹和随后的定量光密度评估以确认Tob-1 mRNA转录本的稳态水平。
阵列分析显示，一种细胞周期调节基因Tob-1在与IL-1β孵育后在两种细胞类型中差异表达。暴露于IL-1β的子宫内膜异位症基质细胞中Tob-1减少了48%，但正常子宫内膜基质细胞中仅减少了16%。重复Northern分析（n = 4）表明，暴露于IL-1β 12小时导致子宫内膜异位症基质细胞中Tob-1 mRNA减少25%±5%。相比之下，在暴露于IL-1β的正常子宫内膜基质细胞中未观察到显著下降（<3%）。
与正常子宫内膜基质细胞相比，细胞周期抑制基因Tob-1在子宫内膜异位症基质细胞中对IL-1β的反应不同。IL-1β下调子宫内膜异位症基质细胞中的Tob-1，但对正常子宫内膜基质细胞没有显著影响。我们的结果表明，IL-1β通过抑制Tob-1促进子宫内膜异位症病变的生长。这些发现首次将IL-1β与子宫内膜异位症种植体来源细胞中的细胞周期基因表达改变联系起来。
