摘要
目的:评价病毒巨噬细胞炎性蛋白ⅡN端肽(NT21MP)是否通过干扰SDF-1α/CXCR4信号抑制人乳腺癌细胞株SKBR3细胞的趋化作用。方法:以RT-PCR和免疫组化检测SKBR3和MCF-7两种人乳腺癌细胞中CXCR4的表达;用细胞转移实验检测在NT21MP存在或缺乏的情况下SDF-1α诱导SKBR3细胞的趋化作用;以Fluo3/AM为细胞内游离钙离子的荧光指示剂,用激光扫描共聚焦显微镜测定NT21MP对SDF-1α诱导SKBR3细胞内游离钙浓度的影响;Western blot分析ERK1/2和FAK蛋白的磷酸化水平变化。结果:相对于MCF-7细胞,SKBR3细胞中CXCR4蛋白表达水平较高;经SDF-1α处理后,SKBR3的迁移能力提高,CXCR4抑制剂AMD3100可有效抑制SKBR3细胞的迁移,细胞经NT21MP预处理后,可剂量依赖性地抑制SKBR3细胞的迁移(P<0.05);NT21MP也可抑制由SDF-1α诱导的细胞内Ca2+峰值(P<0.05),而钙离子浓度升高是SKBR3细胞迁移的重要信号之一;另外,相对于阴性对照组,NT21MP也可下调SDF-1α诱导的SKBR3中信号蛋白ERK1/2和FAK的磷酸化水平(P<0.05)。结论:NT21MP可抑制SDF-1α诱导的SK-BR3细胞的迁移,可能与其上游钙离子释放和ERK1/2及FAK磷酸化阻断信号有关。
AIM： To assess whether NT21MP,the synthetic antagonist 21-mer peptide derived from viral macrophage inflammatory protein Ⅱ inhibits human SKBR3 cells migration by interfering with SDF-1α/CXCR4 signaling.METHODS： The levels of CXCR4 were detected in breast cancer cells SKBR3 and MCF-7 by RT-PCR and immunohistochemistry.The effect of SDF-1α-induced SKBR3 migration（chemotaxis） in the presence and absence of NT21MP was determined using the Boyden chamber migration assay.Intracellular Ca2＋ concentration was measured by fluorometric analysis.Western blot analyses were performed to quantify phosphorylated ERK1/2 and FAK expression levels.RESULTS： The expression of CXCR4 was higher in SKBR3 than MCF-7 cells;SKBR3 migration increased in SDF-1α-treated cells.In contrast,AMD3100,an inhibitor of CXCR4 effectively inhibited SKBR3 migration.SKBR3 migration was decreased when the cells were exposed to NT21MPdose dependently（P0.05）.NT21MP also blocked Ca2＋ influx（P0.05）,an important signal for SKBR3 migration.In addition,NT21MP signiflcantly decreased SDF-1α-induced SKBR3 migration and downregulated SDF-1α-induced expres of phospho-ERK1/2 and phospho-FAK（P0.05）.CONCLUSION： The results showed that NT21MP has an inhibitory effect on SDF-1α-induced SKBR3 migration.The plausible mechanism of action could be upstream blockage of Ca2＋ influx and the downstream reduction of ERK1/2 and FAK signals.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2012年第2期137-140,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助项目(81071848)
安徽省教育厅自然科学研究重点项目(KJ2010A240)
蚌埠市科技计划项目(蚌科[2010]51)
