18 October 2020>: Animal Study
Silencing of Long Non-Coding RNA X Inactive Specific Transcript (Xist) Contributes to Suppression of Bronchopulmonary Dysplasia Induced by Hyperoxia in Newborn Mice via microRNA-101-3p and the transforming growth factor-beta 1 (TGF-β1)/Smad3 AxisWenhao Yuan 1ABCDEF* , Xiaoyan Liu 1ABCDG* , Lingkong Zeng 1BCDFG* , Hanchu Liu 1ABCDEFG , Baohuan Cai 2ABCDEFG , Yanping Huang 1ABDF , Xuwei Tao 1ABCDF , Luxia Mo 1ABDF , Lingxia Zhao 1BCDF , Chunfang Gao 1BCDF
Med Sci Monit 2020; 26:e922424
Figure 3 Xist competitively binds to miR-101-3p to regulate HMGB3. (A) RT-qPCR showed that mRNA expression of HMGB3 in the BPD group was evidently higher than that in the control group. (B) Starbase verified the sequence binding sites between Xist and miR-101-3p and between miR-101-3p and HMGB3. (C) Dual-luciferase reporter gene assay verified the targeting relations between Xist and miR-101-3p and between miR-101-3p and HMGB3. (D) RNA pull-down assay further affirmed the targeting relation between Xist and miR-101-3p. (E) RT-qPCR assessed miR-101-3p expression in mice with different treatment. (F, G) RT-qPCR and Western blot analysis showed that mRNA expression and protein level of HMGB3 in BPD mice were decreased with overexpressed miR-101-3p. Two-way ANOVA was used to assess data in panels A and C, one-way ANOVA was used to assess data in panels D and E. Tukey’s multiple comparisons test was applied for post hoc test. The t test was used for analyzing data in remaining panels. ** p<0.01, ## p<0.01, n=3. Xist – X inactive specific transcript; miR – microRNA; HMGB3 – high-mobility group protein B3; RT-qPCR – reverse transcription-quantitative polymerase chain reaction; BPD – bronchopulmonary dysplasia; ANOVA – analysis of variance.