Wei Ling, Song Xian-Rang, Sun Ju-Jie, Xie Li, Wang Xing-Wu, Lü Li-Yan
Cancer Research Center & Provincial Key Laboratory of Radiation Oncology of Shandong, Shandong Cancer Institute, Jinan, China.
Zhonghua Yi Xue Za Zhi. 2012 Nov 13;92(42):3004-7. doi: 10.3760/cma.j.issn.0376-2491.2012.42.015.
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.
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.
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).
LOX down-regulation reduces invasion and migration potentials of hypoxic human lung cancer cell and potentiates the protein expression of E-cadherin.
观察赖氨酸氧化酶（LOX）下调对缺氧诱导的肺癌NCI-H460细胞侵袭、迁移及上皮-间质转化表型分子E-钙黏蛋白蛋白表达的影响。
在常氧（19%O₂）条件下，使用针对人LOX基因的小干扰RNA（LOX siRNA）转染肺癌细胞。孵育24小时后，将细胞接种于缺氧培养箱（0.5%O₂）中培养24小时。采用实时聚合酶链反应（PCR）检测LOX mRNA表达。通过蛋白质免疫印迹法测定LOX和E-钙黏蛋白的蛋白水平。采用Transwell小室检测细胞的侵袭和迁移能力。
与常氧条件下的NCI-H460细胞（设为1）相比，缺氧使LOX mRNA和蛋白表达水平分别升高至26.04±1.78和5.57±1.27（均P<0.05）。与对照siRNA组（设为1）相比，LOX siRNA转染后LOX mRNA和蛋白表达分别为0.24±0.03和0.29±0.03，细胞侵袭和迁移能力分别为0.57±0.03和0.49±0.02，E-钙黏蛋白的蛋白表达为2.17±0.21（均P<0.05）。
下调LOX可降低缺氧人肺癌细胞的侵袭和迁移能力，并增强E-钙黏蛋白的蛋白表达。
