山东医药2017，Vol.57Issue(3)：16-19,4.DOI:10.3969/j.issn.1002-266X.2017.03.005
SENP1基因沉默的人Ⅱ型肺泡上皮细胞系构建
Establishment of HEPApiCs cell line stably silencing SENP1
摘要
Abstract
Objective To establish human type II alveolar epithelial cells (HEPApiC)that can stably silence SUMO specific protease 1 (SENP 1).Methods We constructed the LV3-SENP1-RNAi lentiviral vector silencing SENP1 gene. HEPApiC cells were divided into the experimental group,control group and empty vector group.The experimental group was infected with LV3-SENP1-RNAi,the empty vector group with empty vector and the control group was not treated.After 72-hour infection,the green fluorescence intensity was observed,the SENP1 mRNA in HEPApiC cells was detected by qRT-PCR,and SENP1 protein was detected by Western blotting.Results After restriction enzyme digestion and sequen-cing,the LV3-SENP 1-RNAi lentiviral vector plasmid was successfully constructed and we got a virion with a high titer. The expression of GFP in HEPApiC cells infected with LV3-SENP1-RNAi was gradually increased with time,indicating that LV3-SENP1-RNAi successfully infected HEPApiC cells.The relative expression of SENP1 mRNA in the experimental group,the control group and the empty vector group was 0.026,0.050,and 0.057;and the relative expression of SENP1 protein was 0.161 ±0.015,0.781 ±0.046 and 0.811 ±0.008,respectively.The expression of SENP1 mRNA and protein in the experimental group was lower than that in the control group and empty vector group,all P <0.05.Conclusion HE-PApiCs cell line stably silencing SENP1 is successfully established.关键词
小泛素相关修饰蛋白质类/人Ⅱ型肺泡上皮细胞/基因沉默/RNA 干扰Key words
small ubiquitin-related modifier proteins/typeⅡalveolar epithelial cells/gene silencing/RNA interfer-ence分类
医药卫生引用本文复制引用
赵许,董文斌,雷小平,李清平,康兰,赵帅,张婵..SENP1基因沉默的人Ⅱ型肺泡上皮细胞系构建[J].山东医药,2017,57(3):16-19,4.基金项目
国家自然科学基金资助项目（81571480）；四川省科技厅科研项目（2014NZ0014）；泸州市政府－泸州医学院联合专项资金资助项目（2013LZLY-J08）。 （）
