Research Paper Volume 13, Issue 17 pp 21743—21757

lncRNA HCG11 suppresses human osteosarcoma growth through upregulating p27 Kip1

HCG11 raises p27 Kip1 expression via binding to miR-942-5p. (A) Predicted binding site of miR-942-5p in HCG11 sequence by StarBase database. (B) MiR-942-5p expression in OS tissues and ANTs was analyzed by qRT-PCR. (C) MiR-942-5p expression in OS cell lines and human osteoblast cell line was analyzed by qRT-PCR. (D) MiR-942-5p level in tumors ≥ 5cm and E) Subcellular co-localization of HCG11 and miR-942-5p was confirmed by RNA in situ hybridization assay. (F) Integration of HCG11 and miR-942-5p was proved by RNA pull-down assay. (G) Specific binding site of miR-942-5p in HCG11 were verified by luciferase assays. (H) Predicted binding site of miR-942-5p in p27 Kip1 3’UTR by StarBase database. (I) Specific binding site of miR-942-5p in p27 Kip1 3’UTR were verified by luciferase assays. (J) Regulation of miR-942-5p on p27 Kip1 protein level was estimated by western blot assay. (K) Regulation of HCG11 via inhibiting miR-942-5p on p27 Kip1 protein level was estimated by western blot assay. **P ***P

Figure 4. HCG11 raises p27 Kip1 expression via binding to miR-942-5p. (A) Predicted binding site of miR-942-5p in HCG11 sequence by StarBase database. (B) MiR-942-5p expression in OS tissues and ANTs was analyzed by qRT-PCR. (C) MiR-942-5p expression in OS cell lines and human osteoblast cell line was analyzed by qRT-PCR. (D) MiR-942-5p level in tumors ≥ 5cm and < 5cm was compared. (E) Subcellular co-localization of HCG11 and miR-942-5p was confirmed by RNA in situ hybridization assay. (F) Integration of HCG11 and miR-942-5p was proved by RNA pull-down assay. (G) Specific binding site of miR-942-5p in HCG11 were verified by luciferase assays. (H) Predicted binding site of miR-942-5p in p27 Kip1 3’UTR by StarBase database. (I) Specific binding site of miR-942-5p in p27 Kip1 3’UTR were verified by luciferase assays. (J) Regulation of miR-942-5p on p27 Kip1 protein level was estimated by western blot assay. (K) Regulation of HCG11 via inhibiting miR-942-5p on p27 Kip1 protein level was estimated by western blot assay. **P < 0.01, ***P < 0.001.