摘要
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.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2012年第42期3004-3007,共4页
National Medical Journal of China