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侵袭迁移和上皮一间质转化表型分子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％0：）肺癌细胞，24h后将细胞置乏氧（0．5％O2）培养箱培养，乏氧24h后采用实时定量PCR检测细胞LOXmRNA表达，Western印迹法评价LOX和E一钙黏素蛋白表达，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LOXmRNA（26．04±1．78）和蛋白（5．574-1．27）表达均显著增加（均P〈0．05）。与乏氧对照siRNA组（设定为1）比较，LOXsiRNA转染后乏氧NCI-H460细胞LOXmRNA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±O．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。

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% Oz ）. After a 24 h incubation, the cells were plated for 24 h in hypoxic incubator （0. 5% Oz ）. 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 transfeetion 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.