Related Experiment Video
Updated: Apr 23, 2026
Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
Effects of shRNA-silenced livin and survivin on lung cancer cell proliferation and apoptosis
Xin Zhao1, Yan Yuan, Zhiqiang Zhang
1Department of Respiratory Medicine, the First Affiliated Hospital of Xinxiang Medical University, Weihui, P.R., China.
Purpose:
To evaluate the effects of short hairpin RNA (shRNA)-mediated silencing of livin and survivin on the proliferation and apoptosis of A549 lung cancer cells.
Methods:
We designed and constructed the eukaryotic expression vectors pSilencer-livin and pSilencer-survivin which contain the Livin and Survivin genes, respectively, and transfected them into liposome-combined lung cancer A549 cells. The cells were then divided into the blank control, plasmid control, Livin, Survivin, and co-transfected groups. Real-time quantitative PCR (qRT-PCR) and Western blot assay were used to determine the mRNA and protein expression levels of Livin and Survivin. The MTT assay was used to evaluate the changes in cell proliferation. The TUNEL assay was used to evaluate the apoptotic rate.
Results:
The shRNA eukaryotic expression vectors of Livin and Survivin were successfully constructed. The mRNA and protein expression of Livin and Survivin were significantly lower in the co-transfected group than in the control groups (p<0.05) . At 48, 60, and 72 hrs after transfection, the cell growth inhibition rate was significantly higher in the co-transfected group than in the single- transfected group (p<0.05) . At 48 hrs after transfection, the apoptotic rate significantly increased (p<0.05) .
Conclusion:
Co-silencing of Livin and Survivin can effectively inhibit the cell proliferation and apoptosis of lung cancer cells.
Insights
Silencing Livin and Survivin using short hairpin RNA (shRNA) effectively reduced lung cancer cell proliferation and increased apoptosis in A549 cells.
Area of Science:
- Molecular biology
- Cancer research
- Gene silencing
Background:
- Livin and Survivin are anti-apoptotic proteins implicated in cancer progression.
- Targeting these proteins offers a potential therapeutic strategy for lung cancer.
Purpose of the Study:
- To investigate the effect of short hairpin RNA (shRNA)-mediated silencing of Livin and Survivin on A549 lung cancer cell proliferation and apoptosis.
Main Methods:
- Construction of eukaryotic expression vectors for Livin and Survivin shRNA.
- Transfection into A549 lung cancer cells.
- Assessment of gene and protein expression using qRT-PCR and Western blot.
- Evaluation of cell proliferation and apoptosis using MTT and TUNEL assays, respectively.
Main Results:
- Successful construction and validation of shRNA vectors for Livin and Survivin.
- Significant reduction in Livin and Survivin mRNA and protein levels in co-transfected cells.
- Marked inhibition of A549 cell proliferation and a significant increase in apoptosis rates post-transfection.
Conclusions:
- Co-silencing of Livin and Survivin demonstrates potent anti-cancer effects.
- This approach effectively inhibits lung cancer cell proliferation and induces apoptosis, suggesting therapeutic potential.
Related Concept Videos
02:15Experimental RNAi
01:22MicroRNAs
01:22MicroRNAs
02:30siRNA - Small Interfering RNAs
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...
01:12Loss of Tumor Suppressor Gene Functions
When the tumor suppressor genes develop mutations or are lost, cells start growing out of control, leading to cancer. However, a single functional copy of the tumor suppressor gene is enough for the cells to maintain their normal functions and cell...
01:20The Retinoblastoma Gene
The first-ever tumor suppressor gene called Rb was identified in retinoblastoma - a rare eye tumor in children. In inherited forms of the disease, a child inherits one defective copy of the Rb gene, which predisposes them to retinoblastoma. However,...