PGE2通过上调CXCR4表达促进SKOV3细胞侵袭的机制研究
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国家自然科学基金资助(81172003)


Prostaglandin E2 promote invasion ability of SKOV3 cell via upregulating CXCR4
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    摘要:

    目的:探讨前列腺素E2(PGE2)上调人卵巢癌SKOV3细胞CXC趋化因子受体4(CXC chemokine receptor 4,CXCR4)的表达从而促进细胞侵袭能力的机制。方法:用PGE2-EP1受体激动剂或抑制剂-CXCR4抑制剂-蛋白激酶C(PKC)抑制剂和钙离子螯合剂处理SKOV3细胞,通过real-time PCR-Western blot-Transwell实验检测CXCR4 mRNA水平-蛋白水平以及细胞的侵袭能力。结果:5 μmol/L PGE2处理SKOV3细胞后,CXCR4 mRNA及蛋白水平与对照组相比分别上升了60.33%-122.88%(P均 < 0.01),细胞的侵袭能力较对照组增强了112.24%(P < 0.01);而经10 μmol/L CXCR4抑制剂AMD3465处理后,细胞的侵袭水平较PGE2处理组下降了54.00%(P < 0.05);5 μmol/L EP1受体激动剂17-PT-PGE2处理SKOV3细胞后,CXCR4 mRNA及蛋白水平与对照组相比分别上升了47.90%(P < 0.01)-85.56%(P < 0.05),细胞的侵袭能力较对照组增强了108.79%(P < 0.01);而10 μmol/L EP1受体抑制剂sc-51322处理后,CXCR4蛋白表达水平较PGE2处理组下降了54.86%(P < 0.05),细胞的侵袭水平较PGE2处理组下降了65.37%(P < 0.05);5 μmol/L PKC抑制剂BIS-1-10 μmol/L钙离子螯合剂BAPTA-AM处理后,CXCR4 蛋白表达水平较17-PT-PGE2处理组分别下降了57.38%-56.14% (P均 < 0.05),细胞的侵袭水平较17-PT-PGE2处理组下降了52.63%-55.26%(P <均0.05)。结论:PGE2可通过激活EP1受体经Gαq/Ca2+/PKC信号转导通路上调卵巢癌SKOV3细胞CXCR4的表达,从而增强肿瘤细胞的侵袭能力。

    Abstract:

    Objective:To investigate the effect of prostaglandin E2 (PGE2) on the invasion ability of ovarian cancer SKOV3 cell via upregulating the receptor of stromal cell-derived factor-1 (chemokine receptor 4,CXCR4). Methods:SKOV3 cells were treated with PGE2,EP1 receptor agonist,CXCR4 antagonist,EP1 receptor antagonist,protein kinase C (PKC) inhibitor and Ca2+ chelating agent. Real-time PCR,Western Blot and Transwell were performed to detect the levels of CXCR4 mRNA and CXCR4 protein as well as the invasion ability in SKOV3 cells. Results:The mRNA level of CXCR4 increased by 60.33% (P <0.01),while the level of CXCR4 protein increased by 122.88% (P < 0.01),and the invasion ability of SKOV3 cells increased by 112.24% (P < 0.01) after treated with 5 μmol/L PGE2. The invasion ability of SKOV3 cells decreased by 54.00% (P < 0.05) after treated with 10 μmol/L CXCR4 antagonist AMD3465 compared with the cells treated with PGE2. When treated with 5 μmol/L EP1 receptor agonist 17-PT-PGE2,the mRNA level of CXCR4 increased by 47.90 % (P <0.01),the level of CXCR4 protein increased 85.56% (P < 0.05),and the invasion ability of SKOV3 cells increased by 108.79% (P < 0.01). The protein level decreased by 54.86% (P < 0.05) and the invasion ability of SKOV3 cells decreased by 65.37% (P < 0.05) after treated with 10 μmol/L EP1 receptor antagonist sc-51322 compared with the cells treated with PGE2. When treated with 5 μmol/L PKC inhibitor BIS-1 and 10 μmol/L Ca2+ chelating agent BAPTA-AM,the protein levels of CXCR4 were decreased by 57.38%(P < 0.05) and 56.14% (P < 0.05) respectively,and the invasion ability of SKOV3 cells decreased by 52.63% (P < 0.05) and 55.26% (P < 0.05) respectively compared with the cells treated with 17-PT-PGE2. Conclusion:PGE2 might up-regulate the expression level of CXCR4 through EP1 receptor which could be partly related to the Gαq/Ca2+/PKC signaling pathway in ovarian cancer SKOV3 cells,and promote the tumor cells invasion.

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汪亦品,冯晓敏,张 海,张 丽,杨沁怡,施 凤,冷 静. PGE2通过上调CXCR4表达促进SKOV3细胞侵袭的机制研究[J].南京医科大学学报(自然科学版),2015,(4):510-516

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  • 收稿日期:2014-12-08
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  • 在线发布日期: 2015-05-05
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