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Updated: May 23, 2026
Evaluating the Effect of SASP Factors on the Proliferation of Cancer Cells Using a Comparative Analysis of Three Distinct Methodologies
Published on: September 19, 2025
Effects of SASH1 on lung cancer cell proliferation, apoptosis, and invasion in vitro
En-guo Chen1, Yanfan Chen, Liang-liang Dong
1Department of Respiratory Medicine, Sir Run Run Shaw Hospital of Zhejiang University, No. 3, East Qingchun Rd, Hangzhou, 310016, China. chenenguo@126.com
Abstract:
The purposes of this study were to investigate the effects of the SASH1 gene on the growth, proliferation, apoptosis, invasiveness, and metastatic potential of lung cancer cells and explore the potential use of SASH1 for the treatment of human lung cancer. The SASH1 gene was cloned into the pcDNA3.1 eukaryotic expression vector, and SASH1 shRNA were designed and constructed. The resulting constructs were transfected into A549 human lung cancer cells, and the changes in the relevant biological characteristics of the cells overexpressing SASH1 and cells with downregulated expression of SASH1 were analyzed using the MTT assay, transwell invasion assay, and flow cytometry. The effects of the SASH1 gene on the expression of cyclin D1, Bcl-2, and MMP-2/9 were also concurrently examined. In the A549 cells from the pcDNA3.1-SASH1 transfected group, cell viability, proliferation, and migration were significantly reduced compared to the control cells (p = 0.039, p = 0.013), and a cell cycle arrest in G1 was observed. The A549 cells transfected with the SASH1 shRNA demonstrated significantly higher cell viabilities, proliferation, and migration compared to the control cells (p = 0.012, p = 0.045). Additionally, the percentage of A549 cells undergoing apoptosis was significantly higher in the pcDNA3.1-SASH1 transfected cells and significantly lower in the SASH1 shRNA transfected cells compared to the control cells (p = 0.010, p = 0.000). The cyclin D1, Bcl-2, and MMP-9/2 protein expression levels were significantly lower in the pcDNA3.1-SASH1-transfected cells and were significantly higher in the SASH1 shRNA-transfected cells than that in the control cells. The SASH1 gene may inhibit A549 cell growth and proliferation as well as promote cellular apoptosis. The overexpression of the SASH1 gene may also be related to the decreased migration of A549 human lung cancer cells.
Insights
The SASH1 gene significantly impacts lung cancer progression. Overexpressing SASH1 inhibits A549 cell growth, proliferation, and migration while promoting apoptosis, suggesting therapeutic potential.
Area of Science:
- Oncology
- Molecular Biology
- Gene Regulation
Background:
- Lung cancer remains a leading cause of cancer-related mortality worldwide.
- Understanding the molecular mechanisms driving lung cancer progression is crucial for developing effective treatments.
- The role of the SASH1 gene in lung cancer biology requires further elucidation.
Purpose of the Study:
- To investigate the effects of the SASH1 gene on lung cancer cell growth, proliferation, apoptosis, invasiveness, and metastasis.
- To explore the potential therapeutic applications of SASH1 in human lung cancer.
Main Methods:
- Cloning of the SASH1 gene into a eukaryotic expression vector (pcDNA3.1).
- Design and construction of SASH1 short hairpin RNA (shRNA).
- Transfection of A549 human lung cancer cells with SASH1 constructs and SASH1 shRNA.
- Analysis of cell viability, proliferation, apoptosis, and migration using MTT assay, transwell invasion assay, and flow cytometry.
- Examination of protein expression levels of cyclin D1, Bcl-2, and MMP-2/9.
Main Results:
- Overexpression of SASH1 in A549 cells significantly reduced cell viability, proliferation, and migration, inducing G1 cell cycle arrest.
- Downregulation of SASH1 using shRNA significantly increased cell viability, proliferation, and migration.
- SASH1 overexpression significantly increased apoptosis, while SASH1 knockdown significantly decreased apoptosis.
- Protein levels of cyclin D1, Bcl-2, and MMP-2/9 were significantly reduced upon SASH1 overexpression and increased upon SASH1 knockdown.
Conclusions:
- The SASH1 gene plays a significant role in inhibiting lung cancer cell growth and proliferation.
- SASH1 promotes apoptosis in lung cancer cells.
- Overexpression of SASH1 is associated with decreased migration of human lung cancer cells, indicating its potential as a therapeutic target.
