摘要
目的研究G蛋白偶联受体相关分选蛋白1(GASP-1)基因干扰对非小细胞肺癌(NSCLC)细胞恶性侵袭和上皮间质转化(EMT)及微管形成的影响。方法以肺癌A549细胞为模型进行研究。将前期试验筛选出的shRNA1用Lipofectamine 2000转染到肺癌A549细胞。分组:对照组、shRNA组和GASP-1-shRNA1组。采用Transwell实验检测细胞侵袭能力,划痕实验检测细胞迁移能力;显微观察上皮间质的形态转化;微管形成实验检测微管的结节数;Western blot检测EMT标志物E-cadherin、N-cadherin、Fibronectin和VEGF蛋白的表达量。结果与对照组相比,GASP-1-shRNA1组中每个视野的细胞侵袭数目减少,划痕愈合率降低,表明干扰GASP-1基因可抑制A549细胞侵袭和迁移。与对照组相比,GASP-1-shRNA1组细胞EMT现象被抑制。微管形成实验中,与对照组相比,GASP-1-shRNA1组细胞微管结节数减少,血管内皮生长因子(VEGF)蛋白表达水平降低。结论GASP-1基因干扰可抑制NSCLC细胞的恶性侵袭、EMT和微管形成。
Objective To study the effects of GDF-associated serum protein-1(GASP-1)gene interference on malignant invasion,epithelial-mesenchymal transition(EMT)and microtubule formation in non-small cell lung carcinoma(NSCLC)cells.Methods Lung cancer A549 cells were used as a model.The shRNA1 screened in the previous experiment was transfected into lung cancer A549 cells with Lipofectamine 2000.Grouping:control group,shRNA group and GASP-1-shRNA1 group.Transwell test was used to detect cell invasion ability,scratch test was used to detect cell migration ability;microscopic observation was used to observe EMT;microtubule formation test was used to detect the number of microtubule nodules;Western blot was used to detect EMT markers E-cadherin,N-Expression of cadherin,Fibronectin and VEGF protein.Results Compared with the control group,the number of cell invasions per field in the GASP-1-shRNA1 group significantly reduced,and the rate of scratch healing significantly reduced,indicating that interference with the GASP-1 gene could significantly inhibit the invasion and migration of A549 cells.Compared with the control group,the EMT in the GASP-1-shRNA1 group was significantly inhibited.In the microtubule formation experiment,compared with the control group,the number of microtubule nodules in the GASP-1-shRNA1 group significantly reduced,and the expression level of vascular endothelial growth factor(VEGF)protein significantly reduced.Conclusion GASP-1 gene interference can significantly inhibit the malignant invasion,EMT and microtubule formation of NSCLC cells.
作者
韩贵良
石岩
焦婷
贾友超
任冠颖
Han Guiliang;Shi Yan;Jiao Ting(Dept of Medical Oncology,Affiliated Hospital of Hebei University,Hebei Key Laboratory of Cancer Radiotherapy and Chemotherapy,Baoding 071000)
出处
《安徽医科大学学报》
CAS
北大核心
2021年第3期396-400,共5页
Acta Universitatis Medicinalis Anhui
基金
2019年保定市科技计划自筹经费项目(编号:1941ZF084)
2016年政府资助省级临床医学优秀人才项目(编号:361007)。
