[Effects of antisense Bmi-1 RNA on the proliferation of lung cancer cell line A549]
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Published on: March 30, 2019
Qi Yu1, Xiu-xiang Meng, Ben Liu
1College of Laboratory Medicine, Dalian Medical University, Dalian 116044, China.
Objective:
To study the effects of antisense Bmi-1 (B cell-specific moloney murine leukemia virus insertion site 1) RNA on the growth, cell cycle and apoptosis of lung cancer cell line A549.
Methods:
Recombinant plasmids carrying antisense Bmi-1 RNA were transfected into A549 cells, which expressed a high level of endogenous Bmi-1. The mRNA level of A549 cell was analyzed by real time quantitative RT-PCR and the protein level was determined using Western blot. MTT growth curve and plate colony forming assay were used to measure the effect of antisense Bmi-1 RNA expression on the growth of A549. Flow cytometry was used to analyze cell cycle and apoptosis.
Results:
Antisense Bmi-1 RNA reduced the Bmi-1 expression at the protein level, but did not alter the mRNA level in A549 cells. Compared with the control cells, A549 cells transfected with antisense Bmi-1 RNA showed a strong inhibition of the cell growth. The number of plate colony formation of the antisense Bmi-1 transfected cells (0.67 +/- 0.50) was less than those of the control (73.0 +/- 4.1) and cells transfected with empty vector (67.0 +/- 4.0, P < 0.01). Transfection of antisense Bmi-1 RNA arrested the A549 cells at G₀/G₁ phase of the cell cycle and did not increase the apoptosis.
Conclusion:
Antisense Bmi-1 RNA expression inhibits A549 cells proliferation, likely through the interference of Bmi-1 leading to an arrest of the proliferating cells at the G₀/G₁ phase.
Insights
Antisense Bmi-1 RNA inhibits lung cancer cell growth by reducing Bmi-1 protein levels and causing cell cycle arrest. This study targeted the B cell-specific moloney murine leukemia virus insertion site 1 (Bmi-1) gene in A549 cells.
Area of Science:
- Oncology
- Molecular Biology
- Gene Regulation
Background:
- Bmi-1 (B cell-specific moloney murine leukemia virus insertion site 1) is implicated in various cancers, including lung cancer.
- Understanding Bmi-1's role is crucial for developing targeted lung cancer therapies.
- The A549 lung cancer cell line expresses high levels of endogenous Bmi-1.
Purpose of the Study:
- To investigate the impact of antisense Bmi-1 RNA on the proliferation, cell cycle, and apoptosis of the A549 lung cancer cell line.
- To determine if targeting Bmi-1 expression can inhibit lung cancer cell growth.
Main Methods:
- A549 cells were transfected with recombinant plasmids encoding antisense Bmi-1 RNA.
- Bmi-1 mRNA and protein levels were quantified using real-time quantitative RT-PCR and Western blot, respectively.
- Cell proliferation was assessed via MTT growth curves and colony formation assays.
- Cell cycle distribution and apoptosis rates were analyzed using flow cytometry.
Main Results:
- Antisense Bmi-1 RNA significantly reduced Bmi-1 protein levels in A549 cells, without affecting mRNA levels.
- Transfection with antisense Bmi-1 RNA markedly inhibited A549 cell proliferation and colony formation compared to controls.
- A significant G₀/G₁ phase cell cycle arrest was observed in cells expressing antisense Bmi-1 RNA.
- No significant increase in apoptosis was detected.
Conclusions:
- Antisense Bmi-1 RNA effectively inhibits the proliferation of A549 lung cancer cells.
- The inhibitory effect is likely mediated by Bmi-1 interference, leading to G₀/G₁ cell cycle arrest.
- Targeting Bmi-1 with antisense RNA presents a potential therapeutic strategy for lung cancer.
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