scc9 cells  (ATCC)

Journal: The Journal of Biological Chemistry

Article Title: Single-cell analysis identifies BASP1 as a driver of drug resistance and cell plasticity in oral squamous cell carcinoma

doi: 10.1016/j.jbc.2025.111126

Figure Lengend Snippet: Single-cell RNA-seq identifies BASP1 expression is elevated in chemoresistant OSCC. A, schematic representation of the experimental workflow for scRNA-seq analysis, of sensitive, early, and late cisplatin-resistant OSCC cells. B, uniform manifold approximation and projection (UMAP) visualization of scRNA-seq data of human OSCC lines (SCC9) presenting with sensitive (SCC9 CisS), early (SCC9 4MCisR) and late cisplatin-resistance (SCC9 8MCisR) patterns. The line shows slingshot pseudotime analysis represents the trajectory of cells transitioning from sensitive to a resistant pattern. C, UMAP projection of scRNA-seq data, highlighting three major cell clusters of (SCC9 CisS), early (SCC9 4MCisR), late (SCC9 8MCisR), and rare transitional clusters (TC) depicted as TC1, TC2. Cells are clustered based on their gene expression profiles and color-coded to distinguish each of them. D, heatmap of top 10 differentially upregulated genes in each identified cell cluster and transitional cluster (TC1 and TC2). E , dot plot showing the average scaled expression of few top differentially regulated genes for annotated cell types and the percentage of cells expressing each gene in major clusters and transitional cluster as depicted in panel D . F, UMAP plot depicting the expression of the BASP1 gene in three major cluster and transitional clusters. The intensity of blue color represents the level of gene expression in individual cells. G, cell lysates from indicated resistant and sensitive OSCC cell lines were subjected to immunoblotting (n = 3) using antibodies against BASP1 and β-Actin. H, relative mRNA expression of BASP1 was analyzed by quantitative real-time PCR (qRT-PCR) in the indicated cells. Data are presented as mean ± SEM (n = 3), ∗ p ≤ 0.05 by 1-way ANOVA. OSCC, oral squamous cell carcinomas.

Article Snippet: SCC9 CisR cell line was lysed in 1% RIPA (CST, Cat No# 9806S) for 30 min on ice to solubilize proteins while maintaining protein-protein interactions.

Techniques: RNA Sequencing, Expressing, Gene Expression, Western Blot, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: The Journal of Biological Chemistry

Article Title: Single-cell analysis identifies BASP1 as a driver of drug resistance and cell plasticity in oral squamous cell carcinoma

doi: 10.1016/j.jbc.2025.111126

Figure Lengend Snippet: BASP1-LIN7A axis regulates β-catenin signalling. A, lysates were isolated from chemoresistant cells stably expressing NtshRNA, LIN7A shRNA and ectopic over expression of LIN7A and subjected to immunoblotting (n = 3) using indicated antibodies. B, lysates were isolated from chemoresistant cells stably expressing NtshRNA, BASP1 shRNA and ectopic over expression of LIN7A (pCMV6 LIN7A) and subjected to immunoblotting (n = 3) using indicated antibodies. C, SCC9 CisR cells were transfected with pCMV6 LIN7A (DDK tagged). Lysates were then isolated, immunoprecipitated with anti-DDK antibody, and immunoblotted with indicated antibodies. D, relative miRNA hsa-miR-501-3P expression levels measured by quantitative PCR. Data are presented as mean ± SEM (n = 3). Statistical significance was determined by one-way ANOVA, ∗∗ p ≤ 0.01. E, lysates were isolated from chemoresistant cells stably expressing NtshRNA, LIN7A shRNA and ectopic over expression LIN7A and subjected to immunoblotting (n = 3) using indicated antibodies. F, lysates were isolated from chemoresistant cells stably expressing NtshRNA, BASP1 shRNA and ectopic over expression LIN7A and subjected to immunoblotting (n = 3) using indicated antibodies. G, chemoresistant cells stably expressing NtshRNA, BASP1 shRNA or LIN7A shRNA were co-transfected with TOPflash and Renilla luciferase reporter vectors. Cells were treated with LiCl (20 mM) for 12 h, and luciferase activity was measured as described in the Methods section. The bar graph shows the relative luciferase activity in each group (n = 3, 2-way ANOVA), ∗ p ≤ 0.05.

Article Snippet: SCC9 CisR cell line was lysed in 1% RIPA (CST, Cat No# 9806S) for 30 min on ice to solubilize proteins while maintaining protein-protein interactions.

Techniques: Isolation, Stable Transfection, Expressing, shRNA, Over Expression, Western Blot, Transfection, Immunoprecipitation, Real-time Polymerase Chain Reaction, Luciferase, Activity Assay

Journal: The Journal of Biological Chemistry

Article Title: Single-cell analysis identifies BASP1 as a driver of drug resistance and cell plasticity in oral squamous cell carcinoma

doi: 10.1016/j.jbc.2025.111126

Figure Lengend Snippet: BASP1 regulates EMT through RAC-alpha serine/threonine-protein kinase (AKT)/β-catenin axis. A, lysates were isolated from chemoresistant cells stably expressing NtshRNA, BASP1 shRNA or ectopic overexpressing BASP1 (pCMV6 BASP1). Immunoblotting (n = 3) was performed using indicated antibodies. B, lysates were isolated from chemoresistant cells stably expressing NtshRNA, BASP1 shRNA, or overexpressing LIN7A (pCMV6 LIN7A) in BASP1 shRNA cells. Immunoblotting (n = 3) was performed using indicated antibodies. C, SCC9 CisR cells were transfected with pCMV6 LIN7A (DDK tagged) or Myr-AKT (constitutively active AKT). Lysates were then isolated, immunoprecipitated with anti-DDK antibody for LIN7A and p-AKT and immunoblotted with indicated antibodies. D, Myr-AKT was overexpressed in cells stably expressing BASP1 shRNA and lysates were isolated from chemoresistant cells and subjected to immunoblotting (n = 3) using indicated antibodies. E, schematic representation of the mechanism by which BASP1 regulates EMT via LIN7A in cisplatin resistant OSCC. BASP1, Brain Abundant Membrane-Attached Signal Protein 1; EMT, epithelial to mesenchymal transition; OSCC, oral squamous cell carcinomas.

Article Snippet: SCC9 CisR cell line was lysed in 1% RIPA (CST, Cat No# 9806S) for 30 min on ice to solubilize proteins while maintaining protein-protein interactions.

Techniques: Isolation, Stable Transfection, Expressing, shRNA, Western Blot, Transfection, Immunoprecipitation, Membrane