CCR4 mediates CCL17 (TARC)-induced migration of human colon cancer cells via RhoA/Rho-kinase signaling (Retracted Article)
CCR4 mediates CCL17 (TARC)-induced migration of human colon cancer cells via RhoA/Rho-kinase signaling (Retracted Article)
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DOI：
10.1007/s00384-013-1712-y
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发表时间：
2013-11-01
影响因子：
2.8
通讯作者：
Thorlacius, Henrik
中科院分区：
文献类型：
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作者：
Al-haidari, Amr A.;Syk, Ingvar;Thorlacius, Henrik
背景 积累的数据表明趋化因子在肿瘤细胞转移中的作用。 CCR4 与血液系统恶性肿瘤有关，最近也与实体瘤有关。在此，我们假设CCR4可能表达并支持结肠癌细胞的迁移。我们使用定量RT-PCR和流式细胞术测定结肠癌细胞系（HT-29）和（AZ-97）上CCR4的mRNA和表面表达。使用 ELISA 和 G-LISA 测定对 CCL17 刺激的结肠癌细胞中的总 RhoA 和活性 RhoA 蛋白水平进行定量。进行迁移测定以评估结肠癌细胞的趋化性。使用增殖测定评估体外肿瘤生长。我们的结果显示结肠癌细胞系 (HT-29) 和肿瘤细胞 (AZ-97) 上 CCR4 的清晰 mRNA 水平和表面表达。 CCR4 配体 CCL17 (TARC) 是结肠癌细胞迁移的有效刺激剂。通过将结肠癌细胞与针对CCR4的抗体或针对CCR4的拮抗剂预温育来抑制这种CCL17诱导的结肠癌细胞迁移。结肠癌细胞中 CCL17 诱导的信号传导表明，CCL17 增加了结肠癌细胞中 RhoA-C mRNA 的形成。我们的结果还发现，CCL17 增加了结肠癌细胞中总 RhoA 和活性 RhoA 蛋白水平。 Rho 激酶抑制剂 Y-27632 消除了 CCL17 诱导的结肠癌细胞趋化性。此外，GGTI-2133对异戊二烯化的抑制显着减少了CCL17引发的结肠癌细胞迁移。我们的新数据首次表明CCL17-CCR4轴可能参与结肠癌细胞的扩散。
Background Accumulating data suggest a role of chemokines in tumor cell metastasis. CCR4 has been implicated in hematologic malignancies and recently also in solid tumors. Herein, we hypothesized that CCR4 might be expressed and support migration of colon cancer cells.We used quantitative RT-PCR and flow cytometry to determine mRNA and surface expression of CCR4 on colon cancer cell lines (HT-29) and (AZ-97). Total RhoA and active RhoA protein levels in CCL17-stimulated colon cancer cells were quantified using ELISA and G-LISA assays. Migration assays were performed to evaluate colon cancer cells chemotaxis. In vitro tumor growth was assessed using proliferation assay.Our results show clear-cut mRNA levels and surface expression of CCR4 on a colon cancer cell line (HT-29) and on tumor cells (AZ-97). CCR4 ligand CCL17 (TARC) was a potent stimulator of colon cancer cell migration. This CCL17-induced colon cancer cell migration was inhibited by pre-incubation of the colon cancer cells with an antibody directed against CCR4 or an antagonist against CCR4. CCL17-induced signaling in colon cancer cells revealed that CCL17 increased mRNA formation of RhoA-C in colon cancer cells. Our results also found that CCL17 increased total RhoA and active RhoA protein levels in colon cancer cells. The Rho-kinase inhibitor Y-27632 abolished CCL17-induced colon cancer cell chemotaxis. In addition, inhibition of isoprenylation by GGTI-2133 markedly reduced colon cancer cell migration triggered by CCL17.Our novel data indicate for the first time that the CCL17-CCR4 axis might be involved in the spread of colon cancer cells.