Possible contribution of aminopeptidase N (APN/CD13) to invasive potential enhanced by interleukin-6 and soluble interleukin-6 receptor in human osteosarcoma cell lines
Possible contribution of aminopeptidase N (APN/CD13) to invasive potential enhanced by interleukin-6 and soluble interleukin-6 receptor in human osteosarcoma cell lines
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DOI：
10.1023/a:1006794617406
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发表时间：
1999-01-01
影响因子：
4
通讯作者：
Roessner, A
中科院分区：
文献类型：
--
作者：
Kido, A;Krueger, S;Roessner, A
本研究旨在阐明氨基肽酶N(APN)的作用，APN是一种依赖锌离子的外肽酶，定位于经促炎症细胞因子处理的人骨肉瘤细胞系细胞表面。我们研究了促炎细胞因子IL-1β、IL-6和肿瘤坏死因子α(TNF-α)以及抗炎细胞因子转化生长因子-β(TGF-β)对APN调节的影响。可溶性白介素6受体(sIL-6R)常与白介素6联合使用，以达到稳定疗效。此外，还检测了骨肉瘤细胞系MG63和HOS的侵袭能力。竞争性RT-PCR和ALA-PNA活性检测显示，IL-6和sIL-6R均能显著增强骨肉瘤细胞株APN的表达和活性。尽管IL-1β显著刺激两种细胞株的APN基因表达，但它只影响MG63细胞的酶活性。然而，肿瘤坏死因子-α和转化生长因子-β对两种细胞株的APN的mRNA表达和酶活性均无影响。在Matrigel侵袭实验中，IL-6和sIL-6R显著上调了这些细胞系的侵袭能力，而其他细胞因子则没有。APN的特异性抑制剂Bestatin可抑制细胞侵袭能力的上调。细胞迁移与APN活性高度相关(r=0.79，P<0.002)。这些发现提示APN有助于IL-6和sIL-6R增强人骨肉瘤的侵袭潜能。
This study aimed at clarifying the role of Aminopeptidase N (APN), a Zn2+-dependent ectopeptidase localized on the cell surface of human osteosarcoma cell lines treated with proinflammatory cytokines. We investigated the proinflammatory cytokines interleukin-1 beta (IL-1 beta), IL-6 and tumor necrosis factor alpha (TNF-alpha) as well as the anti-inflammatory cytokine transforming growth factor beta (TGF-beta) for their influence on APN regulation. Soluble IL-6 receptor (sIL-6R) was always used together with IL-6 to achieve a stable effect. In addition, the invasive potential of the osteosarcoma cell lines MG63 and HOS was examined. Competitive RT-PCR and Ala-pNA activity assays revealed that IL-6 and sIL-6R significantly increased the mRNA expression and activity of APN in both osteosarcoma cell lines. Although IL-1 beta significantly stimulated APN mRNA expression in both cell lines, it influenced the enzyme activity only in MG63. TNF-alpha and TGF-beta, however, had an effect neither on mRNA expression nor on the enzyme activity of APN in both cell lines. In the Matrigel invasion assay, IL-6 and sIL-6R significantly up-regulated the transmigration of these cell lines, whereas other cytokines did not. The up-regulated invasion was inhibited by bestatin, a specific inhibitor of APN. Cellular migration correlated highly with APN activity (r=0.79, P < 0.002). These findings suggest that APN contributes to the invasive potential of human osteosarcomas enhanced by IL-6 and SIL-6R.