乏氧状态干扰Snail对人肺腺癌细胞转移潜能影响及其机制探讨  

Influence of Snail inhibition on invasion of human lung cancer cells under hypoxic conditions and EMT related molecular mechanisms

作　　者：魏玲[1] 孙菊杰[2] 王兴武[1] 吕丽燕[1] 谢丽[1] 宋现让[1] 

机构地区：[1]山东省放射肿瘤学重点实验室山东省肿瘤医院基础研究中心,山东济南250117 [2]山东省放射肿瘤学重点实验室山东省肿瘤医院病理科,山东济南250117

出　　处：《中华肿瘤防治杂志》2013年第23期1803-1806,1811,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：山东省自然科学基金(ZR2010HM083);山东省医药卫生科技发展计划(2011HZ095)

摘　　要：目的:观察乏氧状态下干扰锌指转录因子Snail对人肺腺癌SPCA1细胞侵袭的影响及其上皮-间质转化(epithelial mesenehymal transition,EMT)相关分子机制。方法:应用靶向人Snail基因的小干扰RNA(Snail siRNA)转染常氧(19%O2)肺癌细胞,48h后将细胞置乏氧(0.5%O2)培养箱培养,乏氧24h后采用Real-Time PCR检测细胞Snail mRNA表达,蛋白质印迹法检测Snail和EMT表型分子E-钙黏素蛋白表达,Transwell小室评价细胞侵袭能力。结果:与常氧细胞(设为1)相比,乏氧诱导肺癌SPCA1细胞Snail mRNA(5.31±0.73)和蛋白(4.82±0.67)表达均显著增加,P均<0.05。与乏氧对照siRNA组(设为1)比较,LOX siRNA转染后乏氧SPCA1细胞Snail mRNA和蛋白表达分别为0.26±0.08和0.28±0.03。将常氧细胞侵袭力和E-钙黏素表达设为1,则乏氧细胞侵袭力和E-钙黏素表达分别为1.85±0.13和0.45±0.04,与常氧细胞存在显著差异,P值均<0.05。下调Snail表达后,乏氧细胞侵袭力和E-钙黏素表达分别为0.90±0.08和1.81±0.09,与乏氧对照siRNA组存在显著差异,P值均<0.05。将乏氧对照siRNA细胞侵袭力和E-钙黏素表达设为1,则Snail siRNA组侵袭力和E-钙黏素分别为0.50±0.03和2.04±0.17,P值均<0.05。结论:乏氧状态下干扰Snail降低肺癌细胞侵袭,其机制可能与增加E-钙黏素蛋白表达有关。OBJECTIVE：To observe the influence of zincic transcription Snail interference on invasion of human lung cancer SPCA1 cells under hypoxic conditions and EMT related molecular mechanisms. METHODS： Lung cancer cells under normoxia （19%O2 ） were transfeeted with small interfering RNA targeting human Snail gene （Snail siRNA）. After 48 h,the cells were plated in hypoxic incubator （0.5%O2） for 24 h, Snail mRNA expression was detected by real-time PCR assay. Protein levels of Snail and E-cadherin were determined by western blot assay. Invasion potential was determined by transwell chamber. RESULTS： Compared with normoxic cells （Set to 1）, hypoxia increased the levels of Snail mRNA and protein expression up to 5.31 ±0.73 and 4.82 ±0.67, respectively （both P〈0.05）. Compared with control siRNA group （Set to 1）, Snail mRNA and protein expression a{ter Snail siRNA transfection were 0.26±0.08 and 0.28 ± 0.03, respectively. Cellular invasive capacity and protein expression of E-cadherin under hypoxia were set to 1 ,and they were 1.85±0.13 and 0.45± 0. 04, respectively under hypoxia. There were significant differences among them. Hypoxic invasive capacity and protein expression of E-cadherin in Snail downregulated cells were 0.90±0.08 and 1.81 ±0.09, respectively. Significant differences existed between groups of Snail siRNA and control siRNA （both P〈O. 05）. Cellular invasive capacity and E-cadherin expression in hypoxic control siRNA were set to 1 ,and they were 0.50 ± 0.03 and 2.04±0.17, respectively, in Snail siRNA groups （both P〈0.05）. CONCLUSION： Snail suppression reduces invasion potential of lung cancer ceils under hypoxic conditions, the involved mechanism is associated with potentiated protein expression of E-cadherin.

关 键 词：乏氧 SNAIL 肺肿瘤 侵袭 E-钙黏素 

分 类 号：R734.2[医药卫生—肿瘤]