Volume 35, Issue 251 (12-2025)
J Mazandaran Univ Med Sci 2025, 35(251): 15-29 |
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Rahavi H, Najafi A, Asgarian-omran H, Aflatoonian B, Tehrani M, Farashahi Yazd E. Dual Blockade of PD-L1 and AXL in SKOV3 Cell Line: A Novel Strategy for Immunotherapy in Ovarian Cancer. J Mazandaran Univ Med Sci 2025; 35 (251) :15-29
URL: http://jmums.mazums.ac.ir/article-1-22164-en.html
URL: http://jmums.mazums.ac.ir/article-1-22164-en.html
Hossein Rahavi 
, Ahmad Najafi , Hossein Asgarian-omran , Behrouz Aflatoonian , Mohsen Tehrani , Ehsan Farashahi Yazd
, Ahmad Najafi , Hossein Asgarian-omran , Behrouz Aflatoonian , Mohsen Tehrani , Ehsan Farashahi Yazd
Abstract: (35 Views)
Background and purpose: Ovarian cancer stands out as one of the most lethal forms of gynecological malignancy, marked by elevated mortality rates primarily due to its frequent late-stage diagnosis and the development of therapeutic resistance. This investigation sought to examine a novel combinatorial approach that targets both PD-L1-mediated immune evasion and AXL-driven epithelial–mesenchymal transition (EMT) in SKOV3 ovarian cancer cells.
Materials and methods: For therapeutic intervention, PD-L1 was silenced using sequence-specific siRNA (10 nM; 48-hour transfection), while AXL signaling was inhibited using the selective small-molecule inhibitor R428 (IC₅₀ = 2.9 μM at 48 hours). Experimental groups included scramble siRNA, PD-L1 siRNA, scramble siRNA combined with R428, PD-L1 siRNA combined with R428, R428 monotherapy, and untreated controls. Cell viability was assessed using the MTT assay, and gene expression levels were quantified by quantitative real-time PCR (qRT-PCR).
Results: This investigation demonstrated that SKOV3 cells intrinsically exhibit basal expression of AXL, Vimentin, PD-L1, TGF-β, and MMP-9, indicating a baseline metastatic potential and an inherent capacity for immune evasion. Both single agents (PD-L1 siRNA and R428) exhibited significant anti-proliferative activity (P < 0.0007 and P < 0.0005, respectively). Importantly, their combination revealed a synergistic effect, resulting in significantly greater growth inhibition (P < 0.0001). Gene expression analysis confirmed marked downregulation of PD-L1 (P < 0.0001) and multiple EMT markers following combination treatment, evidenced by a 72% reduction in vimentin, 65% in TGF-β, and 58% in MMP-9 compared to controls.
Conclusion: The findings showed that simultaneous inhibition of the PD-L1 and AXL pathways substantially suppresses cancer progression, resulting in a significant downregulation of EMT-regulating genes.
Materials and methods: For therapeutic intervention, PD-L1 was silenced using sequence-specific siRNA (10 nM; 48-hour transfection), while AXL signaling was inhibited using the selective small-molecule inhibitor R428 (IC₅₀ = 2.9 μM at 48 hours). Experimental groups included scramble siRNA, PD-L1 siRNA, scramble siRNA combined with R428, PD-L1 siRNA combined with R428, R428 monotherapy, and untreated controls. Cell viability was assessed using the MTT assay, and gene expression levels were quantified by quantitative real-time PCR (qRT-PCR).
Results: This investigation demonstrated that SKOV3 cells intrinsically exhibit basal expression of AXL, Vimentin, PD-L1, TGF-β, and MMP-9, indicating a baseline metastatic potential and an inherent capacity for immune evasion. Both single agents (PD-L1 siRNA and R428) exhibited significant anti-proliferative activity (P < 0.0007 and P < 0.0005, respectively). Importantly, their combination revealed a synergistic effect, resulting in significantly greater growth inhibition (P < 0.0001). Gene expression analysis confirmed marked downregulation of PD-L1 (P < 0.0001) and multiple EMT markers following combination treatment, evidenced by a 72% reduction in vimentin, 65% in TGF-β, and 58% in MMP-9 compared to controls.
Conclusion: The findings showed that simultaneous inhibition of the PD-L1 and AXL pathways substantially suppresses cancer progression, resulting in a significant downregulation of EMT-regulating genes.
Type of Study: Research(Original) |
Subject:
Immunology
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