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Functional characterization of APOBEC3B silencing and its downstream effects in vitro. (A) DepMap analysis identifying HPV-negative <t>(SCC-4)</t> and HPV-positive (SCC-154) oral squamous cell carcinoma lines with differential APOBEC3B expression suitable for functional validation. (B) Quantitative PCR analysis showing that APOBEC3B silencing significantly downregulates its downstream targets ASF1B and RPA2, indicating disruption of the APOBEC3B/ASF1B regulatory axis. (C–D) Cisplatin sensitivity, migration, and invasion assays demonstrate that APOBEC3B knockdown reduces cell motility and enhances cisplatin-induced cytotoxicity, as assessed by CellTiter-Glo viability assay (Promega) and Matrigel-based invasion analysis.(E) Tumor spheroid formation assay showing that shAPOBEC3B markedly suppresses both the number and size of tumor spheres in HPV-positive SCC-154 cells, indicating impaired self-renewal capacity and partial re-sensitization of irradiation-resistant cells to treatment.Data represent mean ± SD from three independent experiments; statistical significance was determined by Student's t-test ( P < 0.05). Abbreviations: SD, standard deviation; EMT, epithelial–mesenchymal transition; HPV, human papillomavirus; shRNA, short hairpin RNA.
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Functional characterization of APOBEC3B silencing and its downstream effects in vitro. (A) DepMap analysis identifying HPV-negative <t>(SCC-4)</t> and HPV-positive (SCC-154) oral squamous cell carcinoma lines with differential APOBEC3B expression suitable for functional validation. (B) Quantitative PCR analysis showing that APOBEC3B silencing significantly downregulates its downstream targets ASF1B and RPA2, indicating disruption of the APOBEC3B/ASF1B regulatory axis. (C–D) Cisplatin sensitivity, migration, and invasion assays demonstrate that APOBEC3B knockdown reduces cell motility and enhances cisplatin-induced cytotoxicity, as assessed by CellTiter-Glo viability assay (Promega) and Matrigel-based invasion analysis.(E) Tumor spheroid formation assay showing that shAPOBEC3B markedly suppresses both the number and size of tumor spheres in HPV-positive SCC-154 cells, indicating impaired self-renewal capacity and partial re-sensitization of irradiation-resistant cells to treatment.Data represent mean ± SD from three independent experiments; statistical significance was determined by Student's t-test ( P < 0.05). Abbreviations: SD, standard deviation; EMT, epithelial–mesenchymal transition; HPV, human papillomavirus; shRNA, short hairpin RNA.
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Functional characterization of APOBEC3B silencing and its downstream effects in vitro. (A) DepMap analysis identifying HPV-negative (SCC-4) and HPV-positive (SCC-154) oral squamous cell carcinoma lines with differential APOBEC3B expression suitable for functional validation. (B) Quantitative PCR analysis showing that APOBEC3B silencing significantly downregulates its downstream targets ASF1B and RPA2, indicating disruption of the APOBEC3B/ASF1B regulatory axis. (C–D) Cisplatin sensitivity, migration, and invasion assays demonstrate that APOBEC3B knockdown reduces cell motility and enhances cisplatin-induced cytotoxicity, as assessed by CellTiter-Glo viability assay (Promega) and Matrigel-based invasion analysis.(E) Tumor spheroid formation assay showing that shAPOBEC3B markedly suppresses both the number and size of tumor spheres in HPV-positive SCC-154 cells, indicating impaired self-renewal capacity and partial re-sensitization of irradiation-resistant cells to treatment.Data represent mean ± SD from three independent experiments; statistical significance was determined by Student's t-test ( P < 0.05). Abbreviations: SD, standard deviation; EMT, epithelial–mesenchymal transition; HPV, human papillomavirus; shRNA, short hairpin RNA.

Journal: Journal of Dental Sciences

Article Title: APOBEC3B/ASF1B–TGF-β signaling axis promotes epithelial–mesenchymal transition in HPV-positive oropharyngeal cancer

doi: 10.1016/j.jds.2025.10.039

Figure Lengend Snippet: Functional characterization of APOBEC3B silencing and its downstream effects in vitro. (A) DepMap analysis identifying HPV-negative (SCC-4) and HPV-positive (SCC-154) oral squamous cell carcinoma lines with differential APOBEC3B expression suitable for functional validation. (B) Quantitative PCR analysis showing that APOBEC3B silencing significantly downregulates its downstream targets ASF1B and RPA2, indicating disruption of the APOBEC3B/ASF1B regulatory axis. (C–D) Cisplatin sensitivity, migration, and invasion assays demonstrate that APOBEC3B knockdown reduces cell motility and enhances cisplatin-induced cytotoxicity, as assessed by CellTiter-Glo viability assay (Promega) and Matrigel-based invasion analysis.(E) Tumor spheroid formation assay showing that shAPOBEC3B markedly suppresses both the number and size of tumor spheres in HPV-positive SCC-154 cells, indicating impaired self-renewal capacity and partial re-sensitization of irradiation-resistant cells to treatment.Data represent mean ± SD from three independent experiments; statistical significance was determined by Student's t-test ( P < 0.05). Abbreviations: SD, standard deviation; EMT, epithelial–mesenchymal transition; HPV, human papillomavirus; shRNA, short hairpin RNA.

Article Snippet: Human oropharyngeal squamous cell carcinoma cell lines SCC-4 (HPV-negative) and SCC-154 (HPV-positive) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Functional Assay, In Vitro, Expressing, Biomarker Discovery, Real-time Polymerase Chain Reaction, Disruption, Migration, Knockdown, Viability Assay, Tube Formation Assay, Irradiation, Standard Deviation, shRNA