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作 者:吕懿[1] 张慧芳[1] 付振东[2] 郑金平[1]
机构地区:[1]山西医科大学公共卫生学院卫生毒理学教研室,太原030001 [2]山西医科大学公共卫生学院流行病与卫生统计学教研室,太原030001
出 处:《中国药物与临床》2017年第9期1273-1276,共4页Chinese Remedies & Clinics
基 金:山西医科大学青年基金(02201305);山西省高校产业化项目(20080017)
摘 要:目的探讨Arresten蛋白对宫颈癌Siha细胞迁移的影响及其可能的作用机制。方法采用四甲基偶氮唑盐(MTT)实验、划痕实验和Boyden小室趋化实验检测Arresten蛋白对宫颈癌Siha细胞增殖及迁移能力的影响;通过免疫细胞化学法检测细胞迁移分子标志基质金属蛋白酶-9(MMP-9)的表达水平;利用实时荧光定量聚合酶链反应(PCR)和蛋白质印迹法检测上皮-间质转化(EMT)标志物E-cadherin与N-cadherin的水平变化。结果经3.2μg/ml Arresten蛋白处理的Siha细胞,其增殖能力没有明显改变(P>0.05)。通过对划痕实验中细胞迁移距离和穿越Boyden小室的细胞数目进行分析,发现经3.2μg/ml Arresten蛋白处理的Siha细胞迁移功能受到抑制(P<0.01),细胞内迁移分子标志基质金属蛋白酶(MMP)-9的表达水平也显著降低(P<0.01)。Arresten蛋白可诱导Siha细胞内上皮标志物E-cadherin表达上调(P<0.01),间充质细胞标志物N-cadherin表达下调(P<0.01)。结论 Arresten蛋白对宫颈癌细胞迁移有抑制作用,可能通过抑制EMT途径发挥作用。。Objective To investigate the effect of Arresten protein on the migration of cervical cancer Siha cells and its possible mechanism. Methods The effects of Arresten protein on the proliferation and migration of cer-vical cancer Siha cells were determined by MTT assay, scratch wound healing assay and Boyden chamber chemotaxis assay. The expression of molecular marker of cell migration, matrix metalloproteinase-9 (MMP-9) was determined by immunocytochemistry. The level changes of epithelial-mesenchymal transition markers, E-cadherin and N-cadherin were examined by real-time fluorescence quantitative PCR and Western blotting. Results The proliferation ability of Siha cells treated with 3.2μg/ml Arresten protein did not change significantly (P〉0.05). The cell migration distance in the scratch wound healing assay and the number of cells crossing the Boyden chamber were analyzed. The findings showed that the migration of Siha cells was inhibited after treated with 3.2 μg/ml Arresten protein (P〈0.01), and the expression of the molecular marker of cell migration, MMP-9 significantly decreased (P〈0.01). Arresten protein could up-regulate the expression of epithelial marker E-cadherin in the Siha cells (P〈0.01), and down-regulate the expres-sion of mesenchymal marker N-cadherin (P〈0.01). Conclusion Arresten protein may inhibit the migration of cervical cancer cells and may play a role in inhibiting EMT pathway.
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