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首页> 外文期刊>Cytokine >Identification of major cellular proteins synthesized in response to interleukin-1 and interleukin-6 in human hepatoma HepG2 cells.
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Identification of major cellular proteins synthesized in response to interleukin-1 and interleukin-6 in human hepatoma HepG2 cells.

机译：鉴定人类肝癌HepG2细胞中对白介素1和白介素6合成的主要细胞蛋白。

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摘要

Interleukin-1 (IL-1) and interleukin-6 (IL-6) are principal proinflammatory cytokines inducing the acute phase response of various tissues, including liver. Cultured human hepatoma HepG2 cells were stimulated with IL-1 (10 ng/ml) and IL-6 (10 ng/ml). After 24 h the cells were collected and disrupted by sonication in a lysis buffer containing 8M urea. The extracted cellular proteins were separated by 2D polyacrylamide gel electrophoresis. The gels were stained with Coomassie Brilliant Blue R-250 and the protein spots showing different intensities in comparison to control (unstimulated) cells were excised and subjected to analysis by LC-MS/MS. Alternatively, proteins were stained with SYPRO Ruby. These differentially expressed proteins include seven up-regulated and two down-regulated intracellular proteins of various functions. The identification of three cytokine-responsive proteins was confirmed by biosynthetic labeling with [35S]methionine after incubation of HepG2 cells, and by western blot with specific antisera.
机译：白介素-1（IL-1）和白介素-6（IL-6）是主要的促炎细胞因子，可诱导包括肝脏在内的各种组织的急性期反应。用IL-1（10ng / ml）和IL-6（10ng / ml）刺激培养的人肝癌HepG2细胞。 24小时后，收集细胞，并在含有8M尿素的裂解缓冲液中通过超声处理破坏细胞。提取的细胞蛋白通过2D聚丙烯酰胺凝胶电泳分离。凝胶用考马斯亮蓝R-250染色，切下与对照（未刺激）细胞相比强度不同的蛋白斑点，并通过LC-MS / MS进行分析。另外，蛋白质用SYPRO Ruby染色。这些差异表达的蛋白质包括七个具有各种功能的上调和两个下调的细胞内蛋白。孵育HepG2细胞后，用[35S]蛋氨酸进行生物合成标记，并用特异性抗血清进行蛋白质印迹，从而证实了三种细胞因子应答蛋白的鉴定。

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