Screening And Bioinformatics Analysis Of Differentially Expressed Long Noncoding RNA Involved In Hyperoxia-induced Bronchopulmonary Dysplasia

[Objective]1. We established a hyperoxia-induced bronchopulmonary dysplasia mouse model to obtain differentially expressed lncRNAs involved in BPD using lncRNA array.2. We selected some differentially expressed lncRNAs validated by qRT-PCR to evaluate the reliability of gene chip.3. Bioinformatics analysis of candidate lncRNAs provided experiment base to clarify the exact molecular mechanism that lncRNAs participated in the occurrence and development of BPD.[Methods]1. We exposed C57BL/6J newborn mice to 95% ambient oxygen and successfully made the hyperoxia-induced BPD model identified by lung tissue H&E staining, radial alveolar counts and the expression of BPD biomarkers.2. For lncRNA arraystar screening, total RNA was extracted from BPD lung tissues.3. Differentially expressed lncRNAs that might be involved in BPD were analyzed by bioinformatics analysis and validated by qRT-PCR.4. We analyzed the candidate lncRNAs of secondary structure and their relationship with the adjacent protein-coding genes by using genome browser.[Results]1. Lung tissue pathological biopsy showed larger alveolar diameter, fewer numbers, thicker alveolar interval and different radial alveolar counts statistically (7.1±0.5 VS 4.8±0.3,p<0.05). TGF-βmRNA expression increased 3.2 fold. The mRNA level of Insulin-like growth factor (IGF)-1 increased by 4 fold. In contrast, the mRNA level of vascular endothelial growth factor-a (VEGF-a), decreased by 60%. These pathological pulmonary changes together with BPD biomarker changes were similar to human BPD, which indicated that we successfully made a hyperoxia-induced BPD model.2. Compared with the control group, there are 1769 lncRNAs with differential expression which have more than 2-fold change and significant difference (p<0.05), of which 882 increase more than 2 times and 887 reduce more than 2 times; 140 increase more than 5 times; 71 reduce more than 5 times; 28 increase more than 10 times and 2 reduce more than 10 times.3. We selected 15 lncRNAs for qRT-PCR validation on the basis of larger difference between groups, smaller difference in the same group and larger initial copy numbers. The qPCR results showed statistically significant difference and were consist with microarray data.4.1010001N08Rik was formed antisense overlap with Gata6 by further bioinformatics analysis, so did AK033210 with TNC, which indicated that they may be involved in the occurrence and development of BPD.[Conclusion 11. We successfully made a hyperoxia-induced BPD model.2. A large number of lncRNAs were identified in BPD lung tissues.3. The microarray data was reliable.4.101000lN08Rik was formed antisense overlap with Gata6 by further bioinformatics analysis, so did AK033210 with TNC, which indicated that they may be involved in the occurrence and development of BPD.