Related Experiment Video
Updated: Dec 18, 2025
An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
[MiR-218 Targeting Bmi-1 Inhibits Proliferation of Acute Promyelocytic Leukemia Cells]
Jian-Feng Liu1, Pan He1, De-Feng Pan2
1Department of Pediatrics, The Second Affiliated Hospital, University of South China, Hengyang 421001, Hunan Province, China.
Objective:
To explore the molecular mechanism by which miR-218 targeting Bmi-1 inhibits the proliferation of acute promyelocytic leukemia (APL) cells.
Methods:
APL cell line HL-60 was transfected by miR-218 and RNA-negative control sequences, respectively. The expression of miR-218 in cells was detected by real-time fluorescence quantitative PCR. The effect of transfected miR-218 on the proliferation of APL cells was detected by MTT assay. Cell apoptosis was detected by flow cytometry. The regulation effect of miR-218 on Bmi-1 expression was determined by Western blot. The correlation of miR-218 expressions with Bmi-1 was analyzed by Spearman test. The targeted relationship between miR-218 and Bmi-1 was verified by luciferase assay.
Results:
MTT assay showed that the proliferation of HL-60 cells in vitro was inhibited by high expression miR-218 significantly. Flow cytometry showed that the G1 and G2 phase cells increased while the S phase cells decreased after transfected by miR-218. Western blot showed that the level of Bmi-1 protein in HL-60 cells decreased significantly after transfection of miR-218 (P＜0.05). Spearman correlation analysis showed that the mRNA level of miR-218 negatively correlated with the protein content of Bmi-1 (r=-0.326, P＜0.01). Luciferase assay indicated that Bmi-1 could targeted on miR-218 directly.
Conclusion:
miR-218 can inhibit the proliferation, metastasis and invasion of APL cells, which can be related with the down-regulated of Bmi-1.
Insights
MicroRNA-218 (miR-218) inhibits acute promyelocytic leukemia (APL) cell proliferation by targeting Bmi-1. This study elucidates the molecular mechanism, showing miR-218 downregulates Bmi-1, impacting APL cell growth.
Area of Science:
- Molecular Biology
- Oncology
- Gene Regulation
Background:
- Acute promyelocytic leukemia (APL) is a subtype of leukemia with specific molecular characteristics.
- Understanding the regulatory mechanisms underlying APL cell proliferation is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the molecular mechanism of microRNA-218 (miR-218) in inhibiting acute promyelocytic leukemia (APL) cell proliferation.
- To determine the role of Bmi-1 as a target gene of miR-218 in APL.
Main Methods:
- APL cell line HL-60 was transfected with miR-218 mimics or control sequences.
- Cell proliferation was assessed using MTT assays.
- Cell cycle distribution and apoptosis were analyzed by flow cytometry.
- Bmi-1 protein expression was evaluated by Western blot.
- The direct targeting relationship between miR-218 and Bmi-1 was confirmed using luciferase reporter assays.
Main Results:
- Overexpression of miR-218 significantly inhibited HL-60 cell proliferation.
- miR-218 transfection led to an increase in G1 and G2 phase cells and a decrease in S phase cells.
- Bmi-1 protein levels were significantly downregulated in HL-60 cells following miR-218 transfection.
- A negative correlation was observed between miR-218 expression and Bmi-1 protein levels.
- Luciferase assays confirmed that miR-218 directly targets Bmi-1.
Conclusions:
- miR-218 effectively inhibits the proliferation of APL cells.
- The inhibitory effect of miR-218 on APL cells is mediated through the downregulation of Bmi-1.
- These findings suggest that miR-218 may serve as a potential therapeutic target for APL.
Related Concept Videos
02:23Abnormal Proliferation
02:57Targeted Cancer Therapies
There are several types of targeted therapies against...