Research Paper Volume 13, Issue 10 pp 14258—14276

LncRNA LOC100129620 promotes osteosarcoma progression through regulating CDK6 expression, tumor angiogenesis, and macrophage polarization

LncRNA LOC100129620 regulates the expression of CDK6 through miR-335-3p. (A) Schematic diagram of the wild-type and mutated binding sites of the CDK6 3′-UTR and miR-335-3p. (B) CDK6 mRNA expression in U2OS cells treated with miR-335-3p mimics or inhibitors, as detected by qRT-PCR. (C) Western blot to detect the effect of miR-335-3p on the protein expression of CDK6. (D) Binding of the CDK6 3′-UTR and miR-335-3p, as detected by luciferase reporter gene assay. (E) Western blot to detect the effect of LOC100129620 on the protein expression of CDK6. (F) Western blot to detect the effect of LOC100129620 on the protein expression of CDK6 in U2OS cells treated with miR-335-3p inhibitors. (G) Transwell assay to detect the effect of LOC100129620 on the invasion ability of U2OS cells with CDK6 knockdown. (H) Quantitative analysis of the Transwell assay results to detect the effect of LOC100129620 on the invasion ability of U2OS cells with CDK6 knockdown. (I) Transwell to detect the effect of LOC100129620 on the invasion ability of MG63 cells with CDK6 knockdown. (J) Quantitative analysis of the Transwell assay results to detect the effect of LOC100129620 on the invasion ability of MG63 cells with CDK6 knockdown. Statistical analysis was conducted using Student’s t-test. Values are means ± SD. **P

Figure 6. LncRNA LOC100129620 regulates the expression of CDK6 through miR-335-3p. (A) Schematic diagram of the wild-type and mutated binding sites of the CDK6 3′-UTR and miR-335-3p. (B) CDK6 mRNA expression in U2OS cells treated with miR-335-3p mimics or inhibitors, as detected by qRT-PCR. (C) Western blot to detect the effect of miR-335-3p on the protein expression of CDK6. (D) Binding of the CDK6 3′-UTR and miR-335-3p, as detected by luciferase reporter gene assay. (E) Western blot to detect the effect of LOC100129620 on the protein expression of CDK6. (F) Western blot to detect the effect of LOC100129620 on the protein expression of CDK6 in U2OS cells treated with miR-335-3p inhibitors. (G) Transwell assay to detect the effect of LOC100129620 on the invasion ability of U2OS cells with CDK6 knockdown. (H) Quantitative analysis of the Transwell assay results to detect the effect of LOC100129620 on the invasion ability of U2OS cells with CDK6 knockdown. (I) Transwell to detect the effect of LOC100129620 on the invasion ability of MG63 cells with CDK6 knockdown. (J) Quantitative analysis of the Transwell assay results to detect the effect of LOC100129620 on the invasion ability of MG63 cells with CDK6 knockdown. Statistical analysis was conducted using Student’s t-test. Values are means ± SD. **P < 0.01.