Induction of TNF-alpha, uPA, IL-8 and MCP-1 by doxorubicin in human lung carcinoma cells.
Induction of TNF-alpha, uPA, IL-8 and MCP-1 by doxorubicin in human lung carcinoma cells.
复制标题
阿霉素在人肺癌细胞中诱导 TNF-α、uPA、IL-8 和 MCP-1。
DOI：
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发表时间：
2003
影响因子：
3
通讯作者：
M. Tanimoto
中科院分区：
文献类型：
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作者：
M. Niiya;K. Niiya;T. Kiguchi;M. Shibakura;N. Asaumi;K. Shinagawa;F. Ishimaru;K. Kiura;K. Ikeda;H. Ueoka;M. Tanimoto
目的
我们先前已经证明了阿霉素诱导的尿激酶（uPA）和白细胞介素-8（IL-8）的表达在人H69小细胞肺癌（SCLC）细胞的微阵列技术，使用人类癌症芯片版本2，其中425人的“癌症相关”基因点在板上。微阵列分析还揭示了肿瘤坏死因子-α（TNF-α）的显著诱导，并且多柔比星诱导的巨噬细胞趋化蛋白-1（MCP-1）表达通过RNA酶保护测定证明。我们通过检测阿霉素对其他类型肺癌细胞中TNF-α、uPA、IL-8和MCP-1诱导的影响来扩展研究。
方法
我们研究了阿霉素对12个人肺癌细胞系中TNF-α、uPA、IL-8和MCP-1表达的影响，这些细胞系包括5个SCLC、3个腺癌和4个鳞癌细胞。还研究了它们的受体的表面表达。
结果
TNF-α在三种细胞系H69、SBC-7（SCLC）和PC-9中显著诱导（腺癌），五种细胞系H69、SBC-7、EBC-1中的uPA（鳞状细胞），EBC-2（鳞状细胞）和Sq-1（鳞状细胞），三种细胞系H69、PC-9和EBC-1中的IL-8，以及五种细胞系H69、SBC-3（SCLC）、SBC-7、PC-9和Sq-1中的MCP-1。在H69细胞中，TNF-α抗原水平在阿霉素处理的细胞的条件培养基中增加约5倍，与mRNA水平的增加平行。与uPA和IL-8一样，在2和4 μ M的“亚致死”浓度下观察到最大诱导，在该浓度下，处理后24小时细胞生长受到轻微抑制。此外，细胞不表达包括I型和II型TNF-α受体、uPA受体（uPAR）、C-x-C-趋化因子受体-1（CXCR-1）或C-C-趋化因子受体-2的受体，其分别对应于细胞中由多柔比星诱导的TNF-α、uPA、IL-8和MCP-1，尽管SBC-7细胞表达uPAR，并且EBC-1细胞表达CXCR-1。
结论
在多柔比星刺激后从肿瘤细胞诱导和分泌的TNF-α、uPA、IL-8和MCP-1可以以旁分泌方式激活表达受体的周围细胞，例如嗜中性粒细胞和单核细胞/巨噬细胞。TNF-α是主要的促炎细胞因子，IL-8和MCP-1分别是中性粒细胞和单核细胞/巨噬细胞的主要化学引诱物。此外，uPA激活基质金属蛋白酶9，基质金属蛋白酶9可以截短并激活IL-8。因此，TNF-α、uPA、IL-8和MCP-1的同时诱导可以增强肿瘤和炎症/免疫细胞之间的相互作用，并增加细胞毒性。
PURPOSE
We have previously demonstrated doxorubicin-induced urokinase (uPA) and interleukin-8 (IL-8) expression in human H69 small-cell lung carcinoma (SCLC) cells by a microarray technique using Human Cancer Chip version 2, in which 425 human "cancer-related" genes are spotted on the plates. The microarray analysis also revealed a significant induction of tumor necrosis factor-alpha (TNF-alpha), and doxorubicin-induced macrophage chemoattractant protein-1 (MCP-1) expression was demonstrated by an RNase protection assay. We extended the study by testing the effects of doxorubicin on the induction of TNF-alpha, uPA, IL-8 and MCP-1 in other types of lung carcinoma cells.
METHODS
We investigated the effects of doxorubicin on the expression of TNF-alpha, uPA, IL-8 and MCP-1 in 12 human lung carcinoma cell lines, including five SCLC, three adenocarcinoma and four squamous cell carcinoma cells. The surface expression of their receptors was also investigated.
RESULTS
TNF-alpha was significantly induced in three cell lines, H69, SBC-7 (SCLC) and PC-9 (adenocarcinoma), uPA in five cell lines, H69, SBC-7, EBC-1 (squamous cell), EBC-2 (squamous cell), and Sq-1 (squamous cell), IL-8 in three cell lines, H69, PC-9 and EBC-1, and MCP-1 in five cell lines, H69, SBC-3 (SCLC), SBC-7, PC-9 and Sq-1. In H69 cells, TNF-alpha antigen levels were increased approximately fivefold in the conditioned medium of doxorubicin-treated cells, in parallel with an increase in mRNA levels. As with uPA and IL-8, the maximum induction was observed at the "sublethal" concentrations of 2 and 4 microM at which cell growth was slightly inhibited 24 h after treatment. Furthermore, the cells did not express receptors including types I and II TNF-alpha receptors, uPA receptor (uPAR), C-x-C-chemokine receptor-1 (CXCR-1), or C-C-chemokine receptor-2, corresponding to TNF-alpha, uPA, IL-8 and MCP-1, respectively, that were induced by doxorubicin in the cells, although SBC-7 cells expressed uPAR, and EBC-1 cells expressed CXCR-1.
CONCLUSIONS
TNF-alpha, uPA, IL-8 and MCP-1 induced and secreted from tumor cells upon doxorubicin stimulation may activate surrounding cells expressing the receptors such as neutrophils and monocytes/macrophages in a paracrine fashion. TNF-alpha is a major proinflammatory cytokine, and IL-8 and MCP-1 are major chemoattractants for neutrophils and monocytes/macrophages, respectively. Furthermore, uPA activates matrix metalloproteinase 9 which can truncate and activate IL-8. Thus, the simultaneous induction of TNF-alpha, uPA, IL-8 and MCP-1 may enhance the interaction between tumor and inflammatory/immune cells, and augment cytotoxicity.