CCR4 通过 RhoA/Rho-kinase 信号通路介导 CCL17（TARC）诱导的人结肠癌细胞迁移。
CCR4 mediates CCL17 (TARC)-induced migration of human colon cancer cells via RhoA/Rho-kinase signaling.
机构信息
Department of Clinical Sciences, Malmö, Section of Surgery, Skåne University Hospital, Lund University, 20502, Malmö, Sweden.
出版信息
Int J Colorectal Dis. 2013 Nov;28(11):1479-87. doi: 10.1007/s00384-013-1712-y. Epub 2013 May 7.
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.
METHODS
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.
RESULTS
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.
CONCLUSIONS
Our novel data indicate for the first time that the CCL17-CCR4 axis might be involved in the spread of colon cancer cells.
背景
越来越多的数据表明趋化因子在肿瘤细胞转移中起作用。CCR4 已被牵连到血液恶性肿瘤中，最近也与实体肿瘤有关。在此，我们假设 CCR4 可能在结肠癌细胞的迁移中表达并支持其迁移。
方法
我们使用定量 RT-PCR 和流式细胞术来确定结肠癌细胞系（HT-29）和（AZ-97）中 CCR4 的 mRNA 和表面表达。使用 ELISA 和 G-LISA 测定 CCL17 刺激的结肠癌细胞中总 RhoA 和活性 RhoA 蛋白水平。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。
结果
我们的结果显示结肠癌细胞系（HT-29）和肿瘤细胞（AZ-97）上 CCR4 的明确的 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物。这种 CCL17 诱导的结肠癌细胞迁移可通过用针对 CCR4 的抗体或针对 CCR4 的拮抗剂预先孵育结肠癌细胞来抑制。CCL17 在结肠癌细胞中的信号转导表明 CCL17 增加了结肠癌细胞中 RhoA-C 的 mRNA 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，通过 GGTI-2133 抑制异戊烯化显著降低了 CCL17 触发的结肠癌细胞迁移。
结论
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。
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