Abstract：Objective: To investigate the effect of long non-coding RNA (LncRNA) KCNQ1 opposite strand/antisense transcript 1 (KCNQ1OT1) on the malignant biological behavior of acute myeloid leukemia (AML) cells by targeting miR-324-5p. Methods: The expression levels of LncRNA KCNQ1OT1 and miR-324-5p in human bone marrow stromal cells (HS-5) and AML cell lines (U937, MOLM-13, HL-60) were detected using quantitative real-time PCR (qPCR). RNA immunoprecipitation (RIP), RNA pull-down, and dual-luciferase reporter assays were performed to validate the targeting relationship between LncRNA KCNQ1OT1 and miR-324-5p in U937 cells. U937 cells were divided into five groups: control group, si-KCNQ1OT1 group, miR-324-5p mimics group, si-NC+miR-324-5p-NC group, and si-KCNQ1OT1+miR-324-5p inhibitor group. Cells were transfected with LncRNA KCNQ1OT1 siRNA, miR-324-5p mimics, or corresponding controls. The expression of LncRNA KCNQ1OT1 and miR-324-5p was measured by qPCR. Cell proliferation was assessed using Edu staining, cell invasion and migration were evaluated by Transwell assays, apoptosis was detected by flow cytometry, mitochondrial membrane potential was measured using JC-1 fluorescence staining, and the expression of proliferation-, apoptosis-, and epithelial-mesenchymal transition (EMT)-related proteins was analyzed by Western blot. Results: LncRNA KCNQ1OT1 specifically bound to miR-324-5p in U937 cells. Compared to the control group, the si-KCNQ1OT1 and miR-324-5p mimics groups showed significantly higher miR-324-5p expression (2.21±0.20, 2.30±0.19 vs 0.98±0.13) and apoptosis rates (49.63±2.15%, 54.92±2.67% vs 1.87±0.61%) (P<0.05), while Edu positivity rates (24.14±5.72%, 17.65±4.16% vs 73.52±8.29%), invasion counts (96.83±15.75, 83.50±16.28 vs 327.00±24.56), migration counts (115.33±18.67, 103.50±20.13 vs 383.50±29.25), and mitochondrial membrane potential (0.73±0.21, 0.56±0.18 vs 5.92±0.49) were significantly lower (P<0.05). In the si-KCNQ1OT1+miR-324-5p inhibitor group, miR-324-5p expression (1.02±0.14 vs 2.21±0.20) and apoptosis rates (3.02±0.94% vs 49.63±2.15%) were lower than in the si-KCNQ1OT1 group (P<0.05), while Edu positivity rates (68.93±8.35% vs 24.14±5.72%), invasion counts (309.50±21.90 vs 96.83±15.75), migration counts (368.33±26.40 vs 115.33±18.67), and mitochondrial membrane potential (5.65±0.51 vs 0.73±0.21) were higher (P<0.05). Statistical values were as follows: miR-324-5p expression (F=100.155, P=0.000), apoptosis rate (F=1708.438, P=0.000), Edu positivity rate (F=90.177, P=0.000), invasion count (F=217.555, P=0.000), migration count (F=219.404, P=0.000), and mitochondrial membrane potential (F=311.520, P=0.000). Conclusion: Silencing LncRNA KCNQ1OT1 attenuates the malignant biological behavior of AML cells by upregulating miR-324-5p.
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2025, Vol. 31 
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