Research Paper Volume 11, Issue 16 pp 5975—5991

LINC00511 promotes the malignant phenotype of clear cell renal cell carcinoma by sponging microRNA-625 and thereby increasing cyclin D1 expression

Figure 3. LINC00511 directly sponges miR-625 in ccRCC cells. (A) Nuclear/cytoplasmic fractionation analysis of LINC00511 expression in ccRCC cells. (B) The binding site in LINC00511 for miR-625, as revealed by bioinformatics analysis. The mutant binding sequences in LINC00511 are also shown. (C) A498 and 786-O cells were transfected with the miR-625 mimics or miR-NC. The transfected cells were collected after 48 h incubation and then subjected to RT-qPCR analysis to determine transfection efficiency. *P < 0.05 vs. the miR-NC group. (D) Either LINC00511-Wt or LINC00511-Mut was cotransfected into A498 and 786-O cells with either the miR-625 mimics or miR-NC. After 48 h transfection, the detection of luciferase activity was conducted via a Dual-Luciferase Reporter System. *P < 0.05 vs. the miR-NC group. (E) A RIP assay was conducted to assess the direct interaction between LINC00511 and miR-625. LINC00511 and miR-625 were both immunoprecipitated by the anti-AGO2 antibody from the lysates of A498 and 786-O cells. *P < 0.05 vs. the IgG group. (F) miR-625 expression was quantified in the presence of either si-LINC00511 or si-NC by RT-qPCR. *P < 0.05 vs. the si-NC group. (G) The expression levels of miR-625 in 49 pairs of ccRCC samples and matched adjacent normal renal tissue samples were measured via RT-qPCR. *P < 0.05 vs. normal renal tissues. *P < 0.05 vs. normal renal tissues. (H) An inverse expression correlation between LINC00511 and miR-625 in ccRCC tissue samples was identified by Spearman’s correlation analysis. R2 = 0.3826, P < 0.0001.