Tada M, Diserens A C, Desbaillets I, Jaufeerally R, Hamou M F, de Tribolet N
Department of Neurosurgery, Centre Hospitalier Universitaire de Vaudois, Lansanne, Switzerland.
J Neuroimmunol. 1994 Mar;50(2):187-94. doi: 10.1016/0165-5728(94)90045-0.
We investigated the expression and production of the interleukin-1 receptor antagonist (IL-1ra) in three human glioblastoma cell lines (LN443, LN444, LN859). Reverse transcription-polymerase chain reaction (RT-PCR) demonstrated the expression of IL-1ra mRNA transcripts in the three cell lines. These three cell lines also expressed mRNA for IL-1 alpha, IL-1 beta, as well as IL-1 receptor type I and type II, suggesting the presence of an IL-1 autocrine loop in these cell lines. Northern blot analysis demonstrated that the IL-1ra mRNA expression increased with IL-1 beta or tumor necrosis factor (TNF)-alpha but not with GM-CSF stimulation in both LN443 and LN444 cell lines. PMA stimulation increased the mRNA expression in LN444 but not in LN443 cells. Immunocytochemical staining showed IL-1ra immunoreactivity in these three cell lines. ELISA on culture supernatants demonstrated that the IL-1ra was secreted from the cell lines in agreement with the mRNA expression. RT-PCR with isoform-specific primers showed that both intracellular and secreted forms of IL-1ra were expressed by the three cell lines, with a predominance of the intracellular form. In vivo study with RT-PCR and Northern blot analysis demonstrated IL-1ra mRNA in six out of 12 human glioblastoma and two out of five anaplastic astrocytoma tissues, although the expression level was not high in some cases. Immunohistochemistry demonstrated the presence of IL-1ra within the cytoplasm of tumor cells in six out of 10 glioblastomas in vivo. These results suggest a potential role of IL-1ra in regulation of the IL-1 autocrine loop in glioblastomas.
我们研究了白细胞介素-1受体拮抗剂（IL-1ra）在三种人胶质母细胞瘤细胞系（LN443、LN444、LN859）中的表达及产生情况。逆转录-聚合酶链反应（RT-PCR）证实这三种细胞系中存在IL-1ra mRNA转录本的表达。这三种细胞系还表达IL-1α、IL-1β以及Ⅰ型和Ⅱ型IL-1受体的mRNA，提示这些细胞系中存在IL-1自分泌环。Northern印迹分析表明，在LN443和LN444细胞系中，IL-1ra mRNA表达随IL-1β或肿瘤坏死因子（TNF）-α刺激而增加，但不受粒细胞-巨噬细胞集落刺激因子（GM-CSF）刺激的影响。佛波酯（PMA）刺激使LN444细胞的mRNA表达增加，但对LN443细胞无此作用。免疫细胞化学染色显示这三种细胞系中有IL-1ra免疫反应性。对培养上清液进行酶联免疫吸附测定（ELISA）表明，IL-1ra从细胞系中分泌，与mRNA表达情况一致。用亚型特异性引物进行RT-PCR显示，三种细胞系均表达细胞内和分泌型IL-1ra，且以细胞内形式为主。RT-PCR和Northern印迹分析的体内研究表明，12例人胶质母细胞瘤中有6例以及5例间变性星形细胞瘤中有2例存在IL-1ra mRNA，尽管在某些病例中表达水平不高。免疫组织化学显示，在体内10例胶质母细胞瘤中有6例肿瘤细胞胞质内存在IL-1ra。这些结果提示IL-1ra在胶质母细胞瘤中IL-1自分泌环的调节中可能发挥作用。
