Enhancement of the CXCL12/CXCR4 axis due to acquisition of gemcitabine resistance in pancreatic cancer: effect of CXCR4 antagonists.
Enhancement of the CXCL12/CXCR4 axis due to acquisition of gemcitabine resistance in pancreatic cancer: effect of CXCR4 antagonists.
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DOI：
10.1186/s12885-016-2340-z
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发表时间：
2016-05-12
期刊：
影响因子：
3.8
通讯作者：
Takeyama H
中科院分区：
文献类型：
--
作者：
Morimoto M;Matsuo Y;Koide S;Tsuboi K;Shamoto T;Sato T;Saito K;Takahashi H;Takeyama H
CXCL 12-CXCR 4信号传导轴在恶性肿瘤生物学中的重要性增加，并且这些肽与肿瘤生长、侵袭和转移有关。本研究的目的是检查轴在胰腺癌（PaCa）细胞与吉西他滨耐药（GEM-R）肿瘤中的基质细胞的关系中的重要作用，并确认CXCR 4拮抗剂治疗GEM-R PaCa细胞的有效性。我们利用MIA PaCa-2和AsPC-1细胞建立了两个GEM-R PaCa细胞系。逆转录聚合酶链反应（RT-PCR）检测PaCa细胞CXCR 4 mRNA和成纤维细胞CXCL 12 mRNA的表达。采用免疫细胞化学和ELISA法检测PaCa细胞中CXCR 4蛋白的表达。使用Matrigel侵袭试验和动物研究，我们然后检查了两种CXCR 4拮抗剂AMD 11070和KRH 3955对CXCL 12刺激的GEM-R PaCa细胞的侵袭性和致瘤性的影响。我们发现GEM可显著增强GEM-R PaCa细胞中CXCR 4的表达，但在正常GEM敏感（GEM-S）PaCa细胞中则无此作用。在RT-PCR和ELISA测定中，通过与用GEM处理的GEM-R PaCa细胞共培养，成纤维细胞的CXCL 12的产生和分泌显著增强。在Matrigel侵袭试验中，GEM处理的GEM-R PaCa细胞的侵袭性被成纤维细胞来源的CXCL 12显著激活，并被CXCR 4拮抗剂AMD 11070和KRH 3955显著抑制。在体内，GEM可激活GEM-R PaCa细胞的致瘤性，而加入CXCR 4拮抗剂则可显著抑制其致瘤性。我们的研究结果表明，CXCL 12-CXCR 4信号轴在PaCa细胞对GEM的抗性中起重要作用。在GEM-R PaCa细胞中暴露于GEM显著增强CXCR 4表达，但在GEM-S PaCa细胞中不增强。此外，CXCR 4拮抗剂可以抑制GEM-R PaCa细胞的生长和侵袭。这些药物可能在未来作为GEM-R PaCa的二线化疗药物。本文的在线版本（doi：10.1186/s12885-016-2340-z）包含补充材料，可供授权用户使用。
The CXCL12-CXCR4 signaling axis in malignant tumor biology has increased in importance, and these peptides are implicated in tumor growth, invasion and metastasis. The aim of our study was to examine the important role of the axis in pancreatic cancer (PaCa) cells’ relationship with stromal cells in gemcitabine-resistant (GEM-R) tumors and to confirm the effectiveness of CXCR4 antagonists for the treatment of GEM-R PaCa cells. We established two GEM-R PaCa cell lines using MIA PaCa-2 and AsPC-1 cells. The expression of CXCR4 mRNA in PaCa cells and the expression of CXCL12 mRNA in fibroblasts were examined by reverse transcription polymerase chain reaction (RT-PCR). The expression of CXCR4 protein in PaCa cells was examined by immunosorbent assay (ELISA) and immunocytochemistry. Using Matrigel invasion assays and animal studies, we then examined the effects of two CXCR4 antagonists, AMD11070 and KRH3955, on the invasiveness and tumorigenicity of GEM-R PaCa cells stimulated by CXCL12. We found that the expression of CXCR4 in GEM-R PaCa cells was significantly enhanced by GEM but not in normal GEM-sensitive (GEM-S) PaCa cells. In RT-PCR and ELISA assays, the production and secretion of CXCL12 from fibroblasts was significantly enhanced by co-culturing with GEM-R PaCa cells treated with GEM. In Matrigel invasion assays, the invasiveness of GEM-R PaCa cells treated with GEM was significantly activated by fibroblast-derived CXCL12 and was significantly inhibited by CXCR4 antagonists, AMD11070 and KRH3955. In vivo, the tumorigenicity of GEM-R PaCa cells was activated by GEM, and it was significantly inhibited by the addition with CXCR4 antagonists. Our findings demonstrate that the CXCL12-CXCR4 signaling axis plays an important role in PaCa cells’ resistance to GEM. CXCR4 expression was significantly enhanced by the exposure to GEM in GEM-R PaCa cells but not in GEM-S PaCa cells. Furthermore, CXCR4 antagonists can inhibit the growth and invasion of GEM-R PaCa cells. These agents may be useful as second-line chemotherapy for GEM-R PaCa in the future. The online version of this article (doi:10.1186/s12885-016-2340-z) contains supplementary material, which is available to authorized users.