Research Paper Volume 13, Issue 4 pp 5967—5985

Transfer of lncRNA UCA1 by hUCMSCs-derived exosomes protects against hypoxia/reoxygenation injury through impairing miR-143-targeted degradation of Bcl-2

LncRNA UCA1 could competitively bind to miR-143 to upregulate Bcl-2. (A) Dual luciferase reporter gene assay verified the binding relationship between lncRNA UCA1 and miR-143, and Bcl-2 and miR-143. (B) RNA pull-down verified the binding relationship between lncRNA UCA1 and miR-143, and Bcl-2 and miR-143. (C) Two-color fluorescence in-situ hybridization verified the binding relationship between Bcl-2 and miR-143. The probe of miR-143 was labeled with red fluorescence, the probe of BCL2 was labeled with green fluorescence, and the yellow fluorescence was the overlapping of miR-143 and Bcl2, indicating that miR-143 and BCL2 are directly related. (D) Expression of lncRNA UCA1, miR-143, Bcl-2 and Beclin-1 in cardiomyocytes and myocardial tissue. (E, F) After miR-143 mimic or inhibitor transfection, expression of miR-143, Bcl-2 and Beclin-1 was determined by RT-qPCR; (G) Bcl-2 and Beclin-1 protein levels and MDM2/p53 pathway related protein levels were measured Western blot assay. *p vs. the control group; #p vs. the H/R group. All experiments were repeated 3 times. Data in panels (B and E) were analyzed with one-way ANOVA, and data in panels (A, D, F and G) were analyzed with two-way ANOVA, followed by Tukey’s multiple comparisons test.

Figure 6. LncRNA UCA1 could competitively bind to miR-143 to upregulate Bcl-2. (A) Dual luciferase reporter gene assay verified the binding relationship between lncRNA UCA1 and miR-143, and Bcl-2 and miR-143. (B) RNA pull-down verified the binding relationship between lncRNA UCA1 and miR-143, and Bcl-2 and miR-143. (C) Two-color fluorescence in-situ hybridization verified the binding relationship between Bcl-2 and miR-143. The probe of miR-143 was labeled with red fluorescence, the probe of BCL2 was labeled with green fluorescence, and the yellow fluorescence was the overlapping of miR-143 and Bcl2, indicating that miR-143 and BCL2 are directly related. (D) Expression of lncRNA UCA1, miR-143, Bcl-2 and Beclin-1 in cardiomyocytes and myocardial tissue. (E, F) After miR-143 mimic or inhibitor transfection, expression of miR-143, Bcl-2 and Beclin-1 was determined by RT-qPCR; (G) Bcl-2 and Beclin-1 protein levels and MDM2/p53 pathway related protein levels were measured Western blot assay. *p < 0.05, vs. the control group; #p < 0.05, vs. the H/R group. All experiments were repeated 3 times. Data in panels (B and E) were analyzed with one-way ANOVA, and data in panels (A, D, F and G) were analyzed with two-way ANOVA, followed by Tukey’s multiple comparisons test.