CXCL12/CXCR4 axis triggers the activation of EGF receptor and ERK signaling pathway in CsA-induced proliferation of human trophoblast cells.
CXCL12/CXCR4 axis triggers the activation of EGF receptor and ERK signaling pathway in CsA-induced proliferation of human trophoblast cells.
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CXCL12/CXCR4 轴在 CsA 诱导的人滋养层细胞增殖中触发 EGF 受体和 ERK 信号通路的激活
DOI：
10.1371/journal.pone.0038375
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Li DJ
中科院分区：
文献类型：
--
作者：
Zhao HB;Tang CL;Hou YL;Xue LR;Li MQ;Du MR;Li DJ
前言我们以前的研究已经证明环孢菌素A（CsA）促进人滋养层细胞的增殖。因此，我们进一步研究细胞内信号通路参与CsA诱导的人滋养层细胞增殖。方法采用酶联免疫吸附试验（ELISA）检测CsA对人滋养层细胞CXCL 12分泌的影响。采用免疫荧光和Western blotting方法研究CXCL 12/CXCR 4轴在CsA诱导的人滋养层细胞表皮生长因子受体（EGFR）磷酸化中的作用。采用5-溴-2 ′-脱氧尿苷（BrdU）细胞增殖实验，分析EGFR及其下游细胞外信号调节蛋白激酶（ERK）信号通路在CsA诱导人滋养层细胞增殖中的作用。结果低浓度CsA促进CXCL 12的分泌，重组人CXCL 12促进原代人滋养层细胞和绒癌细胞株JEG-3 EGFR的磷酸化。通过中和抗体或小干扰RNA（siRNA）抑制CXCL 12或CXCR 4可完全阻断CsA诱导的EGFR磷酸化。CsA诱导的人滋养层细胞增殖被EGFR抑制剂AG 1478和ERK抑制剂U 0126有效地消除，但不能被PI 3 K/PKB抑制剂LY 294002消除。CsA促进JEG-3细胞中ERK的活化，其在CXCL 12 siRNA、或CXCR 4 siRNA、或AG 1478存在下被显著废除。结论CsA可能通过CXCL 12/CXCR 4轴促进EGFR活化，EGFR下游ERK信号通路可能参与CsA诱导人滋养层细胞增殖。
Introduction Our previous study has demonstrated Cyclosporin A (CsA) promotes the proliferation of human trophoblast cells. Therefore, we further investigate the intracellular signaling pathway involved in the CsA-induced proliferation of human trophoblast cells. Methods Enzyme-linked immunosorbent assay (ELISA) was performed to evaluate the regulation of CsA on CXCL12 secretion in human trophoblast cells. Immunofluorescence analysis and western blotting analysis were used to investigate the role of CXCL12/CXCR4 axis in the CsA-induced epidermal growth factor receptor (EGFR) phosphorylation in human trophoblast cells. 5-bromo-2′-deoxyuridine (BrdU) cell proliferation assay was performed to analyze the involvement of EGFR and its downstream extracellular signal-regulated protein kinase (ERK) signaling pathway in the CsA-induced proliferation of human trophoblast cells. Results Low concentration of CsA promoted the secretion of CXCL12, and recombinant human CXCL12 promoted the phosphorylation of EGFR in primary human trophoblast cells and choriocarcinoma cell line JEG-3. The inhibition of CXCL12 or CXCR4 by either neutralizing antibodies or small interfering RNA (siRNA) could completely block the CsA-induced EGFR phosphorylation. The CsA-induced proliferation of human trophoblast cells was effectively abrogated by the EGFR inhibitor AG1478 as well as the ERK inhibitor U0126, but not by the PI3K/PKB inhibitor LY294002. CsA promoted the activation of ERK in JEG-3 cells, which was markedly abrogated in the presence of CXCL12 siRNA, or CXCR4 siRNA, or AG1478. Conclusions CsA may promote EGFR activation via CXCL12/CXCR4 axis, and EGFR downstream ERK signaling pathway may be involved in the CsA-induced proliferation of human trophoblast cells.
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