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SEMA4D Knockdown Attenuates β-Catenin-Dependent Tumor Progression in Colorectal Cancer
Mahsa Rezaeepoor1, Golnaz Rashidi1, Mona Pourjafar2
1Department of Immunology, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
Abstract:
Semaphorin 4D (SEMA4D), a protein originally demonstrated to regulate the immune system and axonal growth cone collapse in the developing central nervous system, is overexpressed in various human malignancies, including colorectal cancer (CRC). This investigation was undertaken to examine the effects of SEMA4D silencing on the biological properties of the CRC cell line. SW48 cells were transfected with a siRNA-targeting SEMA4D. The mRNA expression of underlying pro- and antiapoptotic proteins including Bax, Bcl-2, P53, and caspase-3, cancer stem cell (CSC) markers, epithelial-mesenchymal transition (EMT) markers, MMP-2, and MMP-9 was examined using qRT-PCR. Further, the protein expression of E-cadherin and β-catenin was confirmed by Western blot. SW48 cell migration and MMP activity were detected using scratch and zymography analysis, respectively. Finally, the apoptosis rate was assessed via the flowcytometry test. SEMA4D knock-down was associated with a considerable suppression of in vitro cell viability, EMT-related genes, CSC markers, β-catenin signaling pathway, sphere-forming, cell migration, and MMP-2 activity as well as induction of apoptosis. This study identifies the inhibitory effects of SEMA4D gene silencing on tumor progression. Thereby, this might conclude a possible alternative to cancer therapy by targeting several prominent pathways involved in cancer through SEMA4D suppression.
Insights
Silencing Semaphorin 4D (SEMA4D) in colorectal cancer cells suppressed tumor progression by inhibiting cell viability, migration, and epithelial-mesenchymal transition. This suggests SEMA4D as a potential therapeutic target for colorectal cancer.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Semaphorin 4D (SEMA4D) is overexpressed in human malignancies, including colorectal cancer (CRC).
- SEMA4D plays roles in immune regulation and neural development.
- Understanding SEMA4D's role in CRC is crucial for developing new therapies.
Purpose of the Study:
- To investigate the effects of SEMA4D silencing on the biological characteristics of colorectal cancer cells.
- To evaluate the impact of SEMA4D knockdown on key cancer-related pathways and markers.
- To determine the therapeutic potential of targeting SEMA4D in colorectal cancer.
Main Methods:
- SW48 colorectal cancer cells were transfected with siRNA targeting SEMA4D.
- Gene expression (Bax, Bcl-2, P53, caspase-3, CSC markers, EMT markers, MMP-2, MMP-9) was analyzed using qRT-PCR.
- Protein expression (E-cadherin, β-catenin) was assessed via Western blot.
- Cell migration, MMP activity, sphere formation, and apoptosis were evaluated using scratch assays, zymography, and flow cytometry.
Main Results:
- SEMA4D knockdown significantly suppressed in vitro cell viability and sphere-forming capacity.
- Silencing SEMA4D inhibited epithelial-mesenchymal transition (EMT) markers and β-catenin signaling.
- Reduced cell migration and MMP-2 activity were observed, alongside induced apoptosis.
Conclusions:
- SEMA4D gene silencing effectively inhibits colorectal cancer progression in vitro.
- Targeting SEMA4D impacts multiple pathways crucial for tumor growth, including EMT, CSC markers, and MMP activity.
- SEMA4D suppression presents a potential therapeutic strategy for colorectal cancer treatment.
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