赖氨酸氧化酶下调对乏氧肺癌细胞侵袭迁移和E-钙黏素蛋白表达的影响
Effects of lysyl oxidase down-regulation on invasion, migration and E-cadherin protein expression of hypoxic lung cancer cells
目的 观察赖氨酸氧化酶(LOX)下调对乏氧人肺癌细胞NCI-H460侵袭迁移和上皮-间质转化表型分子E-钙黏素蛋白表达的影响.方法 应用针对人LOX基因的小干扰RNA(siRNA)转染常氧(19%O2)肺癌细胞,24 h后将细胞置乏氧(0.5%O2)培养箱培养,乏氧24 h后采用实时定量PCR检测细胞LOX mRNA表达,Western印迹法评价LOX和E-钙黏素蛋白表达,Transwell小室评价细胞侵袭迁移能力.结果 与常氧细胞(设定为1)相比,乏氧诱导肺癌NCI-H460细胞LOX mRNA(26.04±1.78)和蛋白(5.57±1.27)表达均显著增加(均P<0.05).与乏氧对照siRNA组(设定为1)比较,LOX siRNA转染后乏氧NCI-H460细胞LOX mRNA和蛋白表达分别为0.24±0.03和0.29±0.03,细胞侵袭力和迁移力分别为0.57±0.03和0.49±0.02,E-钙黏素蛋白表达为2.17±0.21(均P<0.05).结论 LOX下调可降低乏氧肺癌细胞侵袭迁移,增加E-钙黏素蛋白表达.
更多Objective To observe the effects of lysyl oxidase (LOX) down-regulation on invasion,migration and epithelial-mesenchymal transition phenotype molecule E-cadherin protein expression,induced by hypoxia in lung cancer NCI-H460 cells.Methods Small interfering RNA against human LOX gene (LOX siRNA) was used to transfect lung cancer cells under normoxia (19% O2).After a 24 h incubation,the cells were plated for 24 h in hypoxic incubator (0.5% O2).Real-time polymerase chain reaction (PCR)was performed to detect the LOX mRNA expression.The protein levels of LOX and E-cadherin were determined by Western blot.And invasion and migration capacities were detected by transwell chamber.Results Compared with NCI-H460 cells under normoxia (set to 1),hypoxia increased to the levels of LOX mRNA and protein expression up to 26.04 ± 1.78 and 5.57 ± 1.27 respectively (both P <0.05).Compared with control siRNA group (set to 1),LOX mRNA and protein expression after LOX siRNA transfection were 0.24 ± 0.03 and 0.29 ± 0.03 respectively,cellular invasive and migratory capacities were 0.57 ±0.03 and 0.49 ±0.02 respectively,the protein expression of E-cadherin was 2.17 ±0.21 (all P < 0.05).Conclusion LOX down-regulation reduces invasion and migration potentials of hypoxic human lung cancer cell and potentiates the protein expression of E-cadherin.
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