摘要
目的外源性分化抑制因子3(inhibitor of differentiation 3,Id3)是一种重要的转录调控因子,参与肿瘤发生、细胞增殖和凋亡等过程,文中研究Id3基因表达对人肺腺癌A549细胞迁移性、侵袭性和增殖能力的影响。方法构建人Id3基因和增强型绿色荧光蛋白(enhanced green fluorecence protein,EGFP)真核表达载体pEGFP/Id3,用脂质体转染技术将pEGFP/Id3导入A549细胞,荧光显微镜观察细胞内EGFP的表达情况;RT-PCR分析转染后A549细胞Id3mRNA的表达情况;细胞划痕试验和侵袭小室法检测Id3基因对A549细胞迁移和侵袭能力的影响。平板细胞克隆形成试验测定Id3对A549细胞增殖的影响。结果成功将pEGFP/Id3融合基因转染于A549细胞,荧光显微镜下发出强绿色荧光,pEGFP/Id3转染组的Id3/GAPDH为1.274±0.035,较pEGFP转染组(0.584±0.011)和空白对照组(0.636±0.035)明显上调,差异有统计学意义(P<0.05)。RT-PCR结果显示转染后Id3基因成功表达,细胞划痕实验和侵袭小室法显示转染pEGFP/Id3可明显降低A549细胞的迁移和侵袭能力,pEGFP/Id3转染组48 h的细胞迁移数明显低于空白对照组[(113.75±25.04)个]、[(254.75±41.06)个]和pEGFP转染组[(228.25±26.84)个],差异有统计学意义(P<0.01)。平板克隆试验结果提示外源性Id3可抑制A549细胞增殖,pEGFP/Id3转染组的克隆形成率[(15.67±5.75)%]明显低于空白对照组[(37.83±4.16)%],差异有统计学意义(P<0.05)。结论外源性Id3基因可抑制A549细胞转移及其增殖能力。
Objective The inhibitor of differentiation 3 （Id3）, as an important transcription regular factor, participates in such processes as tumorigenesis, cell proliferation and cell apoptosis. This article aimed to evaluate the effects of Id3 on the migration, inva- sion and proliferation of A549 cells. Methods Recombinant eukaryotic expression vector pEGFP/Id3 was constructed and transfected into A549 cells by the liposome-mediated method. The expression of pEGFP/Id3 in 3.549 cells was analyzed by fluorescence microscopy and RT-PCR. The effects of Id3 on the invasion and migration of A549 cells were detected using the scratch test and invasion chamber model, and that on the proliferation of A549 cells was observed by plate colony formation assay. Results The eukaryotic expression vector pEGFP/Id3 was successfully transfected into the A549 cells. Id3/GAPDH in pEGFP/Id3 was transfection group 1. 274 ± 0.035, which was higher than that in pEGFP transfection group （0.584 ± 0.011 ） and control group （0.636±0. 035）. Id3 was effectively ex- pressed in the A549 cells and significantly reduced their migration and invasion abilities. The transference number of the cells in pEGFP/ Id3 group at 48 hours （ [113.75 ±25.04] ） was lower than that in control group （ [254.75 ±41.06] ） and pEGFP transfection group （ [ 228.25 ± 26.84 % ） （P 〈 0.05）. Plate colony formation assay showed the exogenous Id3 gene transfection evidently inhibited the prolifera- tion of the A549 cells. The success rate of pEGFP/Id3 transfection group （ [ 15.67 ± 5.75 ] % ） was lower significantly than that in control（ [ 37.83 ± 4. 16 ] % ） （P 〈 0.05）. Conclusion The exogenous Id3 ene can inhibit the metastasis and oroliferation of A549 cells.
出处
《医学研究生学报》
CAS
北大核心
2014年第1期15-18,共4页
Journal of Medical Postgraduates
基金
国家自然科学基金(81171652)
关键词
肺腺癌
A549细胞
侵袭
增殖
Lung adenocarcinoma
A549 cell
Invasion
Proliferation
