X-L Bao1, L Zhang, W-P Song. 1. Department of Neonatology, Weinan Maternal and Child Health Hospital, Weinan, China. songwenping12354@163.com.
Abstract
OBJECTIVE: To elucidate the function of long non-coding RNA (lncRNA) SNHG1 in acute myeloid leukemia (AML) and to explore its biological mechanism. PATIENTS AND METHODS: Expression levels of lncRNA SNHG1 and miR-488-5p in AML cell lines pAML and THP-1 were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Proliferative potential and cell cycle progression of pAML and THP-1 cells were detected after SNHG1 knockdown. Potential binding sites of SNHG1 and miR-488-5p were predicted by bioinformatics. Through RNA Binding Protein Immunoprecipitation (RIP) assay and luciferase reporter gene assay, we evaluated the binding condition between SNHG1 and miR-488-5p. MiR-488-5p expression in pAML and THP-1 cells with SNHG1 knockdown was detected by qRT-PCR to further verify their interaction. Subsequently, binding sites of NUP205 and miR-488-5p were predicted. Both mRNA and protein levels of NUP205 in pAML and THP-1 cells were examined. The regulatory effects of overexpressed NUP205 on proliferative potential and cell cycle progression of pAML and THP-1 cells transfected with si-SNHG1 were explored by gain-of-function experiments. RESULTS: LncRNA SNHG1 was highly expressed in pAML and THP-1 cells, while miR-488-5p was lowly expressed. SNHG1 knockdown in pAML and THP-1 cells inhibited proliferative ability and arrested cell cycle in G0/G1 phase. Knockdown of SNHG1 markedly upregulated the miR-488-5p expression in pAML and THP-1 cells. Furthermore, RIP and luciferase reporter gene assay confirmed the binding between SNHG1 and miR-488-5p. NUP205 was highly expressed in pAML and THP-1 cells at both mRNA and protein levels. Moreover, luciferase reporter gene assay indicated that NUP205 could bind to miR-488-5p. More importantly, the overexpression of NUP205 in pAML and THP-1 cells reversed the inhibitory effect of SNHG1 knockdown on proliferative ability and cell cycle progression. CONCLUSIONS: LncRNA SNHG1 promotes the development of AML through miR-488-5p/NUP205 axis.
OBJECTIVE: To elucidate the function of long non-coding RNA (lncRNA) SNHG1 in acute myeloid leukemia (AML) and to explore its biological mechanism. PATIENTS AND METHODS: Expression levels of lncRNA SNHG1 and miR-488-5p in AML cell lines pAML and THP-1 were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Proliferative potential and cell cycle progression of pAML and THP-1 cells were detected after SNHG1 knockdown. Potential binding sites of SNHG1 and miR-488-5p were predicted by bioinformatics. Through RNA Binding Protein Immunoprecipitation (RIP) assay and luciferase reporter gene assay, we evaluated the binding condition between SNHG1 and miR-488-5p. MiR-488-5p expression in pAML and THP-1 cells with SNHG1 knockdown was detected by qRT-PCR to further verify their interaction. Subsequently, binding sites of NUP205 and miR-488-5p were predicted. Both mRNA and protein levels of NUP205 in pAML and THP-1 cells were examined. The regulatory effects of overexpressed NUP205 on proliferative potential and cell cycle progression of pAML and THP-1 cells transfected with si-SNHG1 were explored by gain-of-function experiments. RESULTS: LncRNA SNHG1 was highly expressed in pAML and THP-1 cells, while miR-488-5p was lowly expressed. SNHG1 knockdown in pAML and THP-1 cells inhibited proliferative ability and arrested cell cycle in G0/G1 phase. Knockdown of SNHG1 markedly upregulated the miR-488-5p expression in pAML and THP-1 cells. Furthermore, RIP and luciferase reporter gene assay confirmed the binding between SNHG1 and miR-488-5p. NUP205 was highly expressed in pAML and THP-1 cells at both mRNA and protein levels. Moreover, luciferase reporter gene assay indicated that NUP205 could bind to miR-488-5p. More importantly, the overexpression of NUP205 in pAML and THP-1 cells reversed the inhibitory effect of SNHG1 knockdown on proliferative ability and cell cycle progression. CONCLUSIONS: LncRNA SNHG1 promotes the development of AML through miR-488-5p/NUP205 axis.