3.1 Analysis of uc.375 expression characteristics
We placed newborn mice in a 95% high-oxygen environment for 7 days, and the pathological structure of the mouse lung successfully simulated the characteristics of human BPD. We selected lung tissue samples of the BPD group and the air control group as the test objects, and used lncRNA microarray technology (Mouse lncRNA Microarray V3.0, Arraystar, probes covering 35923 long non-coding RNAs and 24881 coding genes) to screen BPD related lncRNAs. The results suggest that the expression of lncRNA in lung tissue of BPD induced by hyperoxia is quite different from that in normal lung tissue (Figure 1A. Scattered analysis diagram; Figure 1B. Cluster analysis), suggesting that these differences in lncRNA may be the molecular basis for the development of BPD; When analyzing the differentially expressed lncRNAs, it was found that there were 140 upregulated lncRNAs with 5 times or more, and 71 downregulated lncRNAs with 5 times or more.
We also conducted a bioinformatics analysis on uc.375 and identified that uc.375 has a full length of 300bp and is located on human chromosome 14q13 (Figure 1C). It is well-conserved among mammals (Figure 1D and E). There are many transcription factor binding sites in the chromosome region where the uc.375 sequence is located (Figure 1F), suggesting that it may have a potentially powerful transcriptional regulatory effect. We further investigated the expression of uc.375 at different time points in the development of BPD. We exposed newborn mice to 95% oxygen, real time PCR was used to detect the expression of uc.375 in lung tissue at different time points (D0, D1, D3, D5, D7 day) . The results showed that uc.375 decreased gradually with time (Figure 1G). We detected the localization of uc.375 in MLE 12 cells by RNA ISH, and we found that uc.375 existed in the nucleus (Figure 1H).