Expression and Significance of Long non-coding RNA 1010001N08Rik and GATA 6 in Hyperoxia-exposed LLC Cells
Objective:To investigate the expression changes and correlation of long non-coding RNA(lncRNA)1010001N08Rik and GATA6 in hyperoxic LLC cells,and the mechanism of Wnt/β-catenin signaling pathway involved in hyperoxic lung injury.Method:(1)LLC cells were divided into air group.The expression levels of GATA6,Wnt,β-catenin,1010001N08Rik in each group were detected by RT-PCR and the expression levels of related proteins in each group were detected by Western blot.(2)LLC cells were divided into non-interference air group,non-interference hyperoxia group,NC-siRNA interference hyperoxia group and siRNA interference hyperoxia group.The expression levels of GATA6,Wnt,β-catenin,1010001N08Rik in each group were detected by RT-PCR and the expression levels of related proteins in each group were detected by Western blot.The apoptosis was detected by flow cytometry.(3)Data visualization using GraphPad Prism 9.4.1.(4)SPSS 26.0 statistical software was used for data processing.The data of two groups were compared by t test.One-way analysis of variance was used for comparison among multiple groups.(5)Wnt,Fzd2,GATA6 and other related genes were input into string1 1.0 database to construct the PPI network and derive the PPI network diagram.Results:(1)Compared with the air group,the expression of GATA6 mRNA decreased in the hyperoxia exposure group for 24 h and hyperoxia exposure group for 48h,the expression of lncRNA1010001N08Rik mRNA,Wnt mRNA,β-catenin mRNA expression increased.The difference was statistically significant(P＜0.05).(2)Compared with the air group,the expression of GATA6 protein decreased in the hyperoxia exposure group for 24 h and hyperoxia exposure group for 48 h,whlie the expression of Wnt protein,β-catenin protein increased.The difference was statistically significant(P＜0.05).(3)Compared with the non-interference air group,the expression of GATA6 mRNA and protein in the non-interference hyperoxia group and the NC-siRNA interference hyperoxia group decreased,and the expression of 1010001N08Rik mRNA,Wnt mRNA and protein,β-catenin mRNA and protein increased.There was no signifiicant difference between non-interference hyperoxia group and NC-siRNA interference hyperoxia group.Compared with the non-interference hyperoxia group and the NC-siRNA interference hyperoxia group,the expression of GATA6 mRNA and protein in the siRNA interference hyperoxia group increased,and the expression of 1010001N08Rik mRNA,Wnt mRNA and protein,β-catenin mRNA and protein in the siRNA interference hyperoxia group decreased.The difference was statistically significant(P＜0.05).(4)Compared with the non-interference air group,the apoptosis rate of the non-interference hyperoxia group and the NC-siRNA interfered hyperoxia group increased.There was no significant difference between non-interference hyperoxia group and NC-siRNA interference hyperoxia group.Compared with the non-interference hyperoxia group and the NC-siRNA interference hyperoxia group,the apoptosis rate of siRNA interference hyperoxia group decreased.The differences were statistically significant(P＜0.05).(5)According to the protein-protein interaction(PPI)network diagram of Wnt,Fzd2,GATA6 and other related genes derived from the string11.0 database,it can be seen that GATA6 is closely related to Wnt.Conclusion:In LCC cells exposed to hyperoxia,1010001N08Rik may activate Wnt/β-catenin signaling pathway by down-regulating the expression of GATA6,thus promoting the formation process of hyperoxic lung injury.
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