[肿瘤坏死因子α通过p38丝裂原活化蛋白激酶-核因子κB途径诱导人肝癌细胞产生白细胞介素-8]

[TNFα induced IL-8 production through p38 MAPK- NF-kB pathway in human hepatocellular carcinoma cells].

作者信息

Wang Yao-hui, Xia Jing-lin, Wang Wei-min, Yang Bi-wei, Cui Jie-feng, Wang Xiang-dong, Fan Jia

机构信息

Liver Cancer Institute and Zhongshan Hospital, Fudan University, Shanghai 200032, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2011 Dec;19(12):912-6. doi: 10.3760/cma.j.issn.1007-3418.2011.12.008.

DOI:10.3760/cma.j.issn.1007-3418.2011.12.008

PMID:22525504

Abstract

OBJECTIVE

To identify the role of p38 MAPK- NF-kB signaling pathway in TNF-α induced IL-8 production in human hepatocellular carcinoma cells.

METHODS

The concentrations of IL-8 from MHCC-97H cells were measured by an enzyme-linked immunosorbent assay (ELISA). The phosphorylation of p38 MAPK was analyzed by Western blot and immunofluorescence. NF-kB p65 protein nuclear translocation was determined by non-radioactive NF-kB p50 / p65 transcription factor activity kit and immunofluorescence.

RESULTS

The IL-8 production from MHCC-97H cells challenged with TNFa significantly increased in a time-dependent (F = 144.04, P < 0.01) and dose-dependent (F = 364.14, P < 0.01) manners, as compared with those without TNFa challenge. TNFa up-regulated the phosphorylation levels of p38 MAPK and increased the translocation of NF-kB p65 protein into the nucleus, also proved by immunofluorescence staining. p38 MAPK inhibitor (SB203580) could significantly inhibit IL-8 production in a dose-dependent manners (F = 65.47, P < 0.01), and partially inhibited NF-kB p65 nuclear translocation in a dose-dependent manner (F=141.20, P < 0.05).

CONCLUSION

TNF-α could increase the production of IL-8 in MHCC-97H cells and p38 MAPK- NF-kB pathways seem to play a central role in the regulation of IL-8 production.

摘要

目的

确定p38丝裂原活化蛋白激酶（MAPK）-核因子κB（NF-κB）信号通路在肿瘤坏死因子-α（TNF-α）诱导人肝癌细胞白细胞介素-8（IL-8）产生中的作用。

方法

采用酶联免疫吸附测定（ELISA）法检测MHCC-97H细胞中IL-8的浓度。通过蛋白质免疫印迹法和免疫荧光法分析p38 MAPK的磷酸化情况。采用非放射性NF-κB p50/p65转录因子活性试剂盒和免疫荧光法检测NF-κB p65蛋白的核转位。

结果

与未用TNF-α刺激的细胞相比，用TNF-α刺激的MHCC-97H细胞产生的IL-8呈时间依赖性（F = 144.04，P < 0.01）和剂量依赖性（F = 364.14，P < 0.01）显著增加。TNF-α上调p38 MAPK的磷酸化水平，并增加NF-κB p65蛋白向细胞核的转位，免疫荧光染色也证实了这一点。p38 MAPK抑制剂（SB203580）能以剂量依赖性方式显著抑制IL-8的产生（F = 65.47，P < 0.01），并以剂量依赖性方式部分抑制NF-κB p65的核转位（F = 141.20，P < 0.05）。