Establishment of HEPApiCs cell line stably silencing SENP1
ZHAO Xu
DONG Wenbin
LEI Xiaoping
LI Qingping
KANG Lan
ZHAO Shuai
ZHANG Chan
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.
Keywords：small ubiquitin-related modifier proteinstypeⅡalveolar epithelial cellsgene silencingRNA interfer-ence
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 16-19 )