CAJ | 학술논문

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.
목적 관찰뢰안산양화매(LOX)하조대핍양인폐암세포NCI-H460침습천이화상피-간질전화표형분자E-개점소단백표체적영향.방법 응용침대인LOX기인적소간우RNA(siRNA)전염상양(19％O2)폐암세포,24 h후장세포치핍양(0.5％O2)배양상배양,핍양24 h후채용실시정량PCR검측세포LOX mRNA표체,Western인적법평개LOX화E-개점소단백표체,Transwell소실평개세포침습천이능력.결과 여상양세포(설정위1)상비,핍양유도폐암NCI-H460세포LOX mRNA(26.04±1.78)화단백(5.57±1.27)표체균현저증가(균P＜0.05).여핍양대조siRNA조(설정위1)비교,LOX siRNA전염후핍양NCI-H460세포LOX mRNA화단백표체분별위0.24±0.03화0.29±0.03,세포침습력화천이력분별위0.57±0.03화0.49±0.02,E-개점소단백표체위2.17±0.21(균P＜0.05).결론 LOX하조가강저핍양폐암세포침습천이,증가E-개점소단백표체.
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.