Zhang Qingfu, Sun Limei, Yin Liying, Ming Jian, Zhang Siyang, Luo Wenting, Qiu Xueshan
Department of Pathology, the First Affiliated Hospital and College of Basic Medical Sciences of China Medical University, Shenyang, 110001, People's Republic of China,
Tumour Biol. 2013 Oct;34(5):2703-8. doi: 10.1007/s13277-013-0822-z. Epub 2013 May 7.
CCL19/chemokine receptor 7 (CCR7) has been found to be associated with tumor growth, angiogenesis, invasion, and lymph node metastasis. Our previous study demonstrated that CCR7 overexpressed in non-small cell lung cancer (NSCLC) and had close relationship with tumor invasion and lymph node metastasis. However, the molecular mechanism of CCR7 promoting invasion of human NSCLC cells is still unclear. In this study, we demonstrated that human lung adenocarcinoma A549 cells treated with recombinant human CCL19 could obviously upregulate the expression of Sp1 and heparanase at both the mRNA and protein levels. After blockage of CCR7, Sp1 and heparanase expressions were inhibited. Following inhibition of Sp1, heparanase expression was downregulated. The analysis showed the promoter region of heparanase gene containing a number of potential sp1 binding sites (5'-GGGGC-3'). Chromatin immunoprecipitation analysis demonstrated that Sp1 could bind to the heparanase promoter. Cell invasion assays showed that the invasion ability of A549 cells was increased with CCL19 incubation compared to the control cells. These results suggested that CCL19/CCR7 may upregulate the expression of heparanase via Sp1 and contribute to the invasion of A549 cells.
已发现趋化因子CCL19/趋化因子受体7（CCR7）与肿瘤生长、血管生成、侵袭及淋巴结转移相关。我们之前的研究表明，CCR7在非小细胞肺癌（NSCLC）中过表达，且与肿瘤侵袭和淋巴结转移密切相关。然而，CCR7促进人NSCLC细胞侵袭的分子机制仍不清楚。在本研究中，我们证明用重组人CCL19处理人肺腺癌A549细胞后，可在mRNA和蛋白质水平明显上调Sp1和乙酰肝素酶的表达。阻断CCR7后，Sp1和乙酰肝素酶的表达受到抑制。抑制Sp1后，乙酰肝素酶表达下调。分析显示乙酰肝素酶基因的启动子区域含有多个潜在的Sp1结合位点（5'-GGGGC-3'）。染色质免疫沉淀分析表明Sp1可与乙酰肝素酶启动子结合。细胞侵袭试验显示，与对照细胞相比，CCL19孵育可增加A549细胞的侵袭能力。这些结果提示，CCL19/CCR7可能通过Sp1上调乙酰肝素酶的表达，并促进A549细胞的侵袭。
