Research Paper
Circular RNA LPAR3 enhances GPX4 expression by targeting miR-196a-5p to suppress ferroptosis of osteosarcoma cells
Figure 4. CircLPAR3 activates GPX4 expression by targeting miR-196a-5p. (A) The interaction of circLPAR3 with miRNAs was analyzed by RNA-pull down assays in U2OS cells. (B) The prediction analysis using ENCORI database assessed the interaction of miR-196a-5p with circLPAR3 and GPX4 mRNA 3’UTR. (C–E) The analysis was performed in the U2OS and MG63 cells treated with miR-196a-5p mimic. (C) The qPCR measured the miR-196a-5p expression. (D, E) The luciferase reporter gene assays analyzed luciferase activities of circLPAR3 and GPX4 mRNA 3’UTR. (F) The qPCR measured the miR-196a-5p expression in the U2OS and MG63 cells treated with circLPAR3 shRNA. (G) Western blot detected the GPX4 expression in the U2OS and MG63 cells treated with miR-196a-5p mimic. (H) Western blot tested GPX4 expression in the U2OS and MG63 cells co-treated with circLPAR3 shRNA and miR-196a-5p inhibitor. (I) The expression of miR-196a-5p was detected in clinical osteosarcoma samples (n=50) by qPCR. (J) The correlation of miR-196a-5p expression with GPX4 expression was analyzed by qPCR in clinical osteosarcoma samples (n=50). * P < 0.05, ** P < 0.01.