Effect of circ_0000009 on lung adenocarcinoma progression by regulating PDZD2 in a ceRNA- and RBP- dependent manner

Accumulating evidence now demonstrated that circular RNAs (circRNAs) are closely related to the pathogenesis of lung adenocarcinoma (LUAD). Through GEO2R online analysis, we screened hsa_circ_0000009 (circ_0000009) from the GEO database (GSE158695), and its expression in LUAD cancer tissues and cell lines was detected by RT-qPCR. The looping structure of circ_0000009 was tested by RNase R and actinomycin D experiments. The changes of proliferation were tested by CCK-8 or EdU assay. And the changes of apoptosis in A549 and H1299 cells were measured via flow cytometry. The A549 BALB/c tumor model was established to evaluate the influence of circ_0000009 on LUAD cell growth in vivo. In addition, experiments connected with ceRNA direction (mainly including bioinformatics prediction and luciferase reporter assay) and RNA Binding Protein (RBP) direction (mainly including RNA pull-down assay, RIP assay and mRNA stability assay) were further developed to reveal the regulatory mechanism of circ_0000009. The gene and protein levels in this project were assessed by RT-qPCR and western blotting analysis, respectively. The data manifested that circ_0000009 was in low expression in LUAD. The in vitro and in vivo experiments threw light on that overexpression of circ_0000009 dramatically suppressing LUAD tumorigenesis. Mechanistically, circ_0000009 promoted the expression of PDZD2 by sponging miR-154-3p. Furthermore, circ_0000009 stabilized PDZD2 by recruiting IGF2BP2. This study illustrated the mechanism that overexpressing of circ_0000009 suppressed LUAD progression by upregulating PDZD2 expression, providing an original treatment direction for LUAD.