The tetraspanin CD81 mediates the growth and metastases of human osteosarcoma
The tetraspanin CD81 mediates the growth and metastases of human osteosarcoma
复制标题
DOI：
10.1007/s13402-019-00472-w
复制
发表时间：
2019-12-01
影响因子：
6.6
通讯作者：
Kubo, Toshikazu
中科院分区：
文献类型：
--
作者：
Mizoshiri, Naoki;Shirai, Toshiharu;Kubo, Toshikazu
目的CD 81是四跨膜蛋白家族成员。近年来研究表明，CD 81可能参与癌细胞的增殖和转移。方法应用免疫组化方法检测人成骨细胞系hFOB 1.19和人骨肉瘤细胞系Saos 2、MG 63和143 B中CD 81的表达。用siRNA抑制骨肉瘤细胞中CD 81的表达，然后评估细胞的增殖、迁移和侵袭。Western blotting检测Akt、Erk、JNK和p38蛋白的磷酸化状态，并检测MMP-2、MMP-9和MT 1-MMP的表达。此外，我们使用CRISPR/Cas9系统稳定敲除143 B细胞中的CD 81表达，将细胞移植到小鼠体内，并评估这些小鼠与对照组相比的肿瘤形成和肺转移。骨肉瘤细胞系143 B表现出特别高的表达水平。另外，我们发现抑制CD 81后，骨肉瘤细胞的增殖、迁移和侵袭能力降低，Akt和Erk的磷酸化受到抑制。MMP-2、MMP-9和MT 1-MMP的表达水平也受到抑制，其中MMP-9的抑制最大。在体内，我们发现，小鼠移植CD 81敲除143 B细胞表现出显着减少肿瘤形成和肺转移比小鼠在control group.Conclusion根据我们的研究结果，我们得出结论，抑制CD 81抑制细胞内信号转导，并减少骨肉瘤细胞的肿瘤发生和肺转移。
Purpose CD81 is a member of the tetraspanin family of membrane proteins. Recently, it has been shown that CD81 may be involved in cancer cell proliferation and metastasis. As yet, however, there have been few reports on the expression and role of CD81 in osteosarcoma.Methods The expression of CD81 was investigated in human osteoblast cell line hFOB1.19 and in human osteosarcoma cell lines Saos2, MG63 and 143B. The expression of CD81 was inhibited in osteosarcoma cells using siRNA after which cell proliferation, migration and invasion were assessed. We also used Western blotting to investigate the phosphorylation status of Akt, Erk, JNK and p38, and measured the expression ofMMP-2, MMP-9 and MT1-MMP. In addition, we used a CRISPR/Cas9 system to stably knock out CD81 expression in 143B cells, transplanted the cells into mice, and assessed tumor formation and lung metastasis in these mice compared to those in the control group.Results We found that CD81 was expressed in the human osteoblast cell line and in all osteosarcoma cell lines tested. The osteosarcoma cell line 143B exhibited a particularly high level of expression. In addition, we found that osteosarcoma cell proliferation, migration and invasion were decreased after CD81 inhibition, and that the phosphorylation of Akt and Erk was suppressed. Also, the expression levels ofMMP-2, MMP-9 and MT1-MMP were found to be suppressed, withMMP-9 showing the greatest suppression. In vivo, we found that mice transplanted with CD81 knockout 143B cells exhibited significantly less tumor formation and lung metastasis than mice in the control group.Conclusion Based on our findings we conclude that inhibition of CD81 suppresses intracellular signaling and reduces tumorigenesis and lung metastasis in osteosarcoma cells.