沉默HOTAIR通过调控miR-129-5p/EP300轴抑制肾母细胞瘤进展OA

Objective To investigate the effect of long non-coding RNA(lncRNA)HOX transcript antisense RNA(HOTAIR)-based regulation of miR-129-5p/E1A binding protein P300(EP300)axis in nephroblastoma progression.Methods Fifty children with nephro-blastoma who were admitted to the Department of Urology between January 23,2022 and January 23,2024 were enrolled.The levels of HOTAIR,miR-129-5p,and EP300 mRNA expression in nephroblastoma and peritumoral tissues were detected by quantitative real time PCR.The SK-NEP-1 cells were divided as follows:silence control,HOTAIR-silenced,overexpression control,miR-129-5p overexpres-sion,HOTAIR-silenced+inhibitor NC,HOTAIR-silenced+miR-129-5p inhibitor,and control groups.The levels of HOTAIR,miR-129-5p,and EP300 mRNA expression in each group were measured using quantitative real-time PCR.CCK-8 assay,flow cytometry,Transwell assay,wound healing assay and TUNEL staining were used to detect the changes in proliferation,cell cycle,invasion,migration and apoptosis of SK-NEP-1 cells,respectively.Dual-luciferase reporter gene assay was performed to verify the relationships between HOTAIR and miR-129-5p,as well as between miR-129-5p and EP300.The tumor weight and volume were measured in a nude mouse xeno-graft model.Results Compared with peritumoral tissues,in nephroblastoma tissues,the levels of HOTAIR and EP300 mRNA expres-sion were significantly upregulated,whereas miR-129-5p expression was significantly downregulated(all P<0.05).Compared with the control and silence control groups,the HOTAIR-silenced group had reduced HOTAIR and EP300 mRNA expression,optical density(OD)450 value,percentages of cells in S phase and G2/M phase,number of invasive cells,wound-healing rate,and tumor weight and volume in nude mice and increased miR-129-5p expression,percentage of cells in G0/G1 phase,and apoptosis rate(all P<0.05).Compared with the control and overexpression control groups,in the miR-129-5p overexpression group,the EP300 mRNA expression,OD450 value,percen-tages of cells in S and G2/M phases,number of invasive cells,wound-healing rate,and tumor weight and volume in nude mice decreased whereas the miR-129-5p expression,percentage of cells in G0/G1 phase,and apoptosis rate increased(all P<0.05).Compared with the HOTAIR-silenced and HOTAIR-silenced+inhibitor NC groups,in the HOTAIR-silenced+miR-129-5p inhibitor group,EP300 mRNA expression,OD450 value,percentages of cells in S phase and G2/M phase,number of invasive cells,wound-healing rate,and tumor weight and volume in nude mice increased whereas the miR-129-5p expression,percentage of cells in G0/G1 phase,and apoptosis rate decreased(all P<0.05).HOTAIR directly targeted miR-129-5p,and miR-129-5p further directly targeted EP300.Conclusion Silencing HOTAIR potentially inhibits proliferation,invasion,and migration;induces cell-cycle arrest at the G0/G1 phase;promotes apoptosis in SK-NEP-1 cells;and suppresses tumor growth in nude mice via regulation of the miR-129-5p/EP300 axis.