摘要
目的:探讨低氧对人肺腺癌A549细胞来源的A549多细胞球体上皮-间质转化(epithelial-mesenchymal transition,EMT)促转移机制的影响。方法:采用无血清培养(serum-freemedium,SFM)的方法培养A549细胞,诱导A549多细胞球体形成;随后将亲本A549细胞和A549多细胞球体在低氧环境下培养24h,分别检测2种细胞中EMT标志物E-钙黏着素(E-cadherin)和波形蛋白(vimentin)的表达水平,并与它们在常氧条件下的表达情况进行比较。结果:在常氧和低氧条件下,A549多细胞球体中E-cadherinmRNA和蛋白的表达水平均较亲本A549细胞降低,而vimentin的表达水平升高;与常氧条件相比,低氧培养的亲本A549细胞及A549多细胞球体中E-cadherinmRNA和蛋白的表达水平均下调,且以A549多细胞球体下调更为明显,而vimentin的表达水平则升高。结论:A549多细胞球体比亲本A549细胞具有更强的转移能力,低氧条件下A549多细胞球体比常氧条件下的A549多细胞球体具有更强的转移能力。
Objective: To investigate the influence of hypoxia on metastasis induced by epithelialmesenchymal transition of human lung adenocarcinoma A549 cell spheres. Methods: A549 cell spheres were formed through serum-free culture. The expression levels of E-cadherin and vimentin in the parental A549 cells and A549 sphere cells under hypoxia condition for 24 h were examined by immunofluorescence, Western blotting, and reverse transcription (RT)-PCR, and compared with the expressions levels under normoxia condition. Results: Under hypoxia and normoxia conditions, the expression levels of E-cadherin mRNA and protein in the A549 cells spheres were lower than those in the parental A549 cells, but the expression levels of vimentin mRNA and protein in the A549 cell spheres were increased. As compared with the normoxic condition, the expression levels of E-cadherin mRNA and protein in the parental A549 cells and the A549 cell spheres were all down-regulated under hypoxic condition, and especially for the parental A549 cells, this decline was much more obvious; while the expression level of vimentin was increased. Conclusion: A549 cell spheres have a stronger ability of metastasis than the parental A549 cells. This ability of metastasis is also stronger in the A549 cell spheres under hypoxic condition than that in the parental A549 cells under normoxia.
出处
《肿瘤》
CAS
CSCD
北大核心
2012年第3期182-188,共7页
Tumor
基金
江西省教育厅科学技术研究基金资助项目(编号:GJJ10345)