human cscc cell line a431  (ATCC)

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Flowchart overview of single-cell sequencing in human skin of actinic keratosis (AK), squamous cell carcinoma in situ (SCCIS), and cutaneous squamous cell carcinoma (cSCC) patients. ( B ) Hematoxylin and eosin (H&E) staining of skin biopsies from representative AK (100× and 250×), SCCIS (50× and 250×), and cSCC (50× and 250×). ( C ) Uniform manifold approximation and projection (UMAP) plot of human normal skin labeled by cell type and patient, respectively. ( D ) Heatmap showing gene expression signatures of each cell type. ( E ) Violin plot displaying the expression of representative genes to identify subpopulations for each cell type. ( F ) Representative gene ontology (GO) terms of signature genes in different cell subpopulations. The color keys from yellow to red indicate the range of p-value.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Sequencing, In Situ, Staining, Labeling, Gene Expression, Expressing

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Violin plots showing the expression levels of ALDH3A1 and IGDBP2 in actinic keratosis (AK), squamous cell carcinoma in situ (SCCIS), and matched normal samples (Wilcoxon test, p_val_adj <0.05). ( B ) Violin plots showing the expression levels of ALDH3A1 and IGDBP2 in cutaneous squamous cell carcinoma (cSCC) and matched normal samples. ( C ) The effect of UVB on ALDH3A1 expression in HaCaT cells. Left, the cells were collected at 24 hr, 48 hr, 72 hr, 96 hr, and 120 hr after UVB irradiation with a UVB dose of 10 mJ/cm 2 per 24 hr; right, the cells were collected at 24 hr after UVB irradiation with UVB doses of 10 mJ/cm 2 , 20 mJ/cm 2 , 30 mJ/cm 2 , 40 mJ/cm 2 , and 50 mJ/cm 2 . ( D ) The changes of cell proliferation after IGFBP2 overexpression in HaCaT and A431 cells. ( E ) The changes of cell invasion after IGFBP2 overexpression in HaCaT and A431 cells. * p<0.05; **p<0.01; ***p<0.001; ns, not significant.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Expressing, In Situ, Irradiation, Over Expression

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Representative gene ontology (GO) terms for genes with specific expression in basal cell of SCCIS, actinic keratosis (AK), and normal samples in P2 (upper) and up-regulated differentially expressed genes (DEGs) from AK versus normal, SCCIS versus normal, SCCIS versus AK, respectively (lower). ( B ) Heatmap showing copy number variation (CNV) levels of all keratinocytes from AK and SCCIS samples in P2. The keratinocytes from normal sample in P2 were defined as references. ( C ) Uniform manifold approximation and projection (UMAP) of subgroups generated from basal cells in SCCIS sample showing basal cells with higher CNV level enriched in one subgroup; red dot representing basal cells with higher CNV level (cnv.score>81.5). ( D ) DEGs detected between Basal-SCCIS-tumor and Basal-SCCIS-normal. ( E ) Representative enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) and GO terms in up-regulated genes (avg_log 2 FC >0.58 and p_val_adj <0.05). ( F ) Violin plots showing the expression level of representative DNA damage response marker genes in Basal-SCCIS-tumor and Basal-SCCIS-normal subgroups. ( G ) Chord plot showing the top up-regulated genes included in representative GO terms. ( H ) Violin plots showing the expression level of major members in HSP family across Basal-SCCIS-tumor and Basal-SCCIS-normal subgroups (Wilcoxon test, p_val_adj <0.05). ( I ) Violin plots showing the expression level of MAGEA4 and ITGA6 in Basal-SCCIS-tumor and Basal-SCCIS-normal subgroups (Wilcoxon test, p_val_adj <0.05). ( J ) Immunohistochemistry staining of MAGEA4 and ITGA6 in human skin of normal, SCCIS, and cutaneous squamous cell carcinoma (cSCC) samples.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Expressing, Generated, Marker, Immunohistochemistry, Staining

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Violin plots showing the different expression levels of candidate genes across all types of keratinocytes in P2, cutaneous squamous cell carcinoma (cSCC), and normal groups (Wilcoxon test, p_val_adj <0.05). ( B ) The statistical results of immunohistochemistry staining of MAGEA4 in different stages of cSCC including SCCIS, well-differentiated cSCC (WD cSCC), and moderately differentiated/poorly differentiated cSCC (MD/PD cSCC), respectively. ( C ) The statistical results of immunohistochemistry staining of ITGA6 in different stages of cSCC including SCCIS, WD cSCC, and MD/PD.cSCC, respectively.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Expressing, Immunohistochemistry, Staining

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) The mRNA expression of MAGEA4 and ITGA6 in human immortalized keratinocytes (HaCaT) and cutaneous squamous cell carcinoma (cSCC) cell lines (A431, SCL-I, SCL-II). *p<0.05; **p<0.01; ***p<0.001; ns, not significant. ( B ) Effect of small interfering RNA (siRNA) on the expression of MAGEA4 in A432 and ITGA6 in A431, SCL-I, and SCL-II determined by quantitative real-time PCR (qRT-PCR). ( C ) Functional experiment of MAGEA4 in A432. Upper, left, effect of MAGEA4 cSCC cell proliferation by CCK-8 proliferation in A431; upper, right, the effect of MAGEA4 on cSCC cell apoptosis was measured by staining with Annexin V-FITC/PI, followed by FACS analysis. Lower, left, the scratch experiment showed that MAGEA4 knockdown resulted in a shorter vertical migration distance compared with the control group after 72 hr; lower, right, transwell assay showed that the invasion abilities of the si-MAGEA4 groups significantly decreased compared with the si-NC group. **p<0.01. ( D ) Effect of ITGA6 on cSCC cell proliferation by CCK-8 proliferation assay in A431, SCL-I, and SCL-II. *p<0.05; **p<0.01. ( E ) The effect of ITGA6 on cSCC cell apoptosis. **p<0.01. ( F ) ITGA6 knockdown resulted in a shorter vertical migration distance compared with the control group after 72 hr. *p<0.05; **p<0.01. ( G ) The invasion abilities of the si-ITGA6 groups significantly decreased compared with the si-NC group. *p<0.05; **p<0.01.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Expressing, Small Interfering RNA, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Functional Assay, CCK-8 Assay, Staining, Knockdown, Migration, Control, Transwell Assay, Proliferation Assay

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Uniform manifold approximation and projection (UMAP) of all cells from cSCC patients labeled by sample and cell type, respectively. ( B ) Cell proportion of keratinocytes in cSCC and normal groups. ( C ) Expression of basal, Pro KC, and differentiated genes in all keratinocytes of cSCC and normal groups. ( D ) Left, immunohistochemical staining of LGALS1, IFITM3, and FTH1 in normal skin (200×), well-differentiated cSCC (WD cSCC) (50× and 250×), and moderately differentiated/poorly differentiated cSCC (MD/PD cSCC) (50× and 250×). Scale bar, 200 μm and 50 μm. Right, the immunoreactivity score (IRS) analyses of LGALS1, IFITM3, and FTH1 in normal skin, WD cSCC, and MD/PD cSCC. n=15 for each group. *p<0.05; **p<0.01; ***p<0.001; ns, not significant. ( E ) The mRNA expression of LGALS1, IFITM3, and FTH1 in human immortalized keratinocytes (HaCaT) and cSCC cell lines (A431, SCL-I, SCL-II). *p<0.05; **p<0.01; ***p<0.001; ns, not significant.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Labeling, Expressing, Immunohistochemical staining, Staining

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: The changes of cell proportion and significance test in all cutaneous squamous cell carcinoma (cSCC) samples and patient-matched normal skin samples (t test, p<0.05).

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques:

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Heatmap showing CNV levels of all keratinocytes from all cSCC samples. The keratinocytes from all patient-matched normal samples were defined as references. ( B ) Left, heatmap showing CNV levels of all keratinocytes from well-differentiated cSCC (WD cSCC) sample; right, the proportion of tumor and normal cells in WD cSCC sample defined by cnv.cut (probs = 0.99). ( C ) Left, heatmap showing CNV levels of all keratinocytes from poorly differentiated cSCC (PD cSCC) sample; right, the proportion of tumor and normal cells in PD cSCC sample defined by cnv.cut (probs = 0.99). ( D ) Left, heatmap showing CNV levels of all keratinocytes from moderately differentiated cSCC (MD cSCC) sample; right, the proportion of tumor and normal cells in MD cSCC sample defined by cnv.cut (probs = 0.99).

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques:

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) The enriched gene ontology (GO) terms of up-regulated differentially expressed genes (DEGs) in Basal1, Basal2, Pro KC, Follicular2, Spinous1 and Spinous2 between cSCC and normal groups. ( B ) Violin plots showing the different expression levels of candidate genes in cSCC and normal groups (Wilcoxon test, p_val_adj <0.05). ( C ) Left, immunohistochemical staining of BST2 and SAT1 in cSCC in normal skin (200×), well-differentiated cSCC (WD cSCC) (50× and 250×) and moderately differentiated/poorly differentiated cSCC (MD/PD cSCC) (50× and 250×). Scale bar, 200 μm and 50 μm. Right, the immunoreactivity score (IRS) analyses of BST2 and SAT1 in normal skin, WD cSCC and MD/PD cSCC. n=15 for each group. *p<0.05; **p<0.01; ***p<0.001; ns, not significant. ( D ) The mRNA expression of BST2 and SAT1 in human immortalized keratinocytes (HaCaT) and cSCC cell lines (A431, SCL-I, SCL-II). *p<0.05; **p<0.01; ***p<0.001; ns, not significant.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Effect of small interfering RNA (siRNA) on the expression of LGALS1, IFITM3, and FTH1 in A431, SCL-I, and SCL-II determined by quantitative real-time PCR (qRT-PCR). ( B ) Effect of LGALS1, IFITM3, and FTH1 on cSCC cell proliferation. The CCK-8 proliferation assay demonstrated a significant decrease in the proliferation of the si-LGALS1, si-IFITM3, and si-FTH1 groups compared with the si-NC group. *p<0.05; **p<0.01; ***p<0.001; ns, not significant. ( C ) The effect of LGALS1, IFITM3, and FTH1 on cSCC cell apoptosis. Significant increase in the apoptosis of the si-LGALS1, si-IFITM3, and si-FTH1 groups compared with the si-NC group. *p<0.05; **p<0.01; ***p<0.001; ns, not significant. ( D ) The scratch experiment showed that LGALS1 and IFITM3 knockdown resulted in a shorter vertical migration distance compared with the control group after 72 hr, while there was no significant change in the si-FTH1 group. *p<0.05; **p<0.01; ***p<0.001; ns, not significant. ( E ) Transwell assay showed that the invasion abilities of the si-LGALS1, si-IFITM3, and si-FTH1 groups significantly decreased compared with the si-NC group. *p<0.05; **p<0.01; ***p<0.001.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Small Interfering RNA, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, CCK-8 Assay, Proliferation Assay, Knockdown, Migration, Control, Transwell Assay

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Effect of small interfering RNA (siRNA) on the expression of BST2 and SAT1 in A431, SCL-I, and SCL-II determined by quantitative real-time PCR (qRT-PCR). ( B ) Effect of BST2 and SAT1 on cutaneous squamous cell carcinoma (cSCC) cell proliferation by CCK-8 proliferation assay in A431, SCL-I, and SCL-II. **p<0.01. ( C ) The effect of BST2 and SAT1 on cSCC cell apoptosis. **p<0.01. ( D ) BST2 and SAT1 knockdown resulted in a shorter vertical migration distance compared with the control group after 72 hr. *p<0.05; **p<0.01. ( E ) The invasion abilities of the si-ITGA6 groups significantly decreased compared with the si-NC group. *p<0.05; **p<0.01.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Small Interfering RNA, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, CCK-8 Assay, Proliferation Assay, Knockdown, Migration, Control

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Identification of TME cell subpopulations based on their marker genes, including T cells, dendritic (DC) cells, and stromal cells. ( B ) Cell proportion of cell subpopulations in T cells, DC cells, and stromal cells, respectively. ( C ) The monotonically changed differentially expressed genes (DEGs) in TME cells in cSCC samples of all stages (Wilcoxon test, p_val_adj <0.05). ( D ) Comparison of cell interactions among the different clinical stages of cSCC (Wilcoxon test, ****p<0.0001). ( E ) Comparison of total incoming path weights vs total outgoing path weights across all cell populations in three cSCC samples, respectively (Wilcoxon test, p<0.1). ( F ) Circle plot showing the inferred intercellular communication signaling strength network of MHC-II pathway in three cSCC samples, respectively (Wilcoxon test, p<0.1).

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Marker, Comparison

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Uniform manifold approximation and projection (UMAP) of subpopulations generated from fibroblasts in all cutaneous squamous cell carcinoma (cSCC) samples. ( B ) Feature plots showing the expression of signature genes for . ( C ) Feature plots showing the expression of signature genes for Fib3. ( D ) Feature plots showing the expression of signature genes for Fib4.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Generated, Expressing

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Comparison of cell interactions between poorly differentiated cSCC and matched normal samples (Wilcoxon test, ****p<0.0001). ( B ) Circle plot showing the inferred intercellular communication signaling strength network of MHC-II, LAMININ, and TNF pathways in poorly differentiated cSCC and matched normal samples were predicted by CellPhoneDB approach (Wilcoxon test, p<0.1).

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: Comparison

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) Left, uniform manifold approximation and projection (UMAP) of single-cell RNA sequencing (scRNA-seq) dataset from poorly differentiated cSCC (PD cSCC) sample labeled by cell type; right, UMAP plot of single-cell ATAC sequencing (scATAC-seq) dataset from PD cSCC sample labeled by cell type after integration and label transfer with scRNA-seq data. ( B ) Bar plot of annotated differentially accessible region (DAR) location for each type. ( C ) Fragment coverage (frequency of Tn5 insertion) around the DAR on the gene KRT5 and CD83. ( D ) Left, heatmap of average chromVAR motif activity for each cell type. The color scale represents a z-score scaled by row. Right, UMAP plot displaying chromVAR motif activity, gene activity, and gene expression of TP63 (upper) and FOSL1 (lower). The color scale for each plot represents a normalized log-fold-change for the respectively assay. ( E ) Cell-specific mean chromVAR motif activity from the JASPAR database was plotted against cell-specific average expression for the corresponding transcription factor for all cell types and transcription factors. ( F ) Mean chromVAR activity was plotted against average expression for TP63 (left) and FOSL1 (right). Significant correlation was assessed with Pearson’s product moment correlation coefficient using the cor.test function in R.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: RNA Sequencing, Labeling, Sequencing, Activity Assay, Gene Expression, Expressing

Journal: eLife

Article Title: Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma

doi: 10.7554/eLife.85270

Figure Lengend Snippet: ( A ) The attribution plot showing the similarity between Basal-SCCIS-tumor/normal with the cell populations defined by Ji et al. ( B ) The classification heatmap showing the similarity between Basal-SCCIS-tumor/normal with the cell populations defined by Ji et al. ( C ) The attribution plot showing the similarity between basal cells in cutaneous squamous cell carcinoma (cSCC) samples with the cell populations defined by Ji et al. ( D ) The classification heatmap showing the similarity between basal cells in cSCC samples with the cell populations defined by Ji et al. SCCIS, squamous cell carcinoma in situ.

Article Snippet: Human immortalized epidermal keratinocytes cell line (HaCaT cells) and human cSCC cell line A431, SCL-I, and SCL-II ( ; ) were used in this study and were obtained from the American Type Culture Collection (ATCC) and Free University of Berlin.

Techniques: In Situ

Journal: Scientific Reports

Article Title: Eribulin inhibits growth of cutaneous squamous cell carcinoma cell lines and a novel patient-derived xenograft

doi: 10.1038/s41598-023-35811-3

Figure Lengend Snippet: Eribulin suppresses tumor growth in cSCC cell lines in vitro. The sensitivity to eribulin of two cSCC cells (A431 and DJM-1) and of normal human dermal fibroblasts (NHDFs) was determined by ATP assay. 2000 cells per well were seeded in 96-well plates for 24 h. Following that, the cells were treated with eribulin at various concentrations (0.001, 0.1, 0.25, 0.5, 1, 5, 10 nM). After 72 h, cell viability was assessed using CellTiter-Glo ® 2.0 assay (Promega). The x-axis indicates the log value of eribulin concentration; the y-axis indicates the relative cell viability (normalized by no treatment). Data are presented as mean ± SD from triplicate experiments.

Article Snippet: The human cSCC A431 cell line and normal human primary dermal fibroblasts (NHDFs) were purchased from the American Type Culture Collection (ATCC).

Techniques: In Vitro, ATP Assay, Concentration Assay

Journal: Scientific Reports

Article Title: Eribulin inhibits growth of cutaneous squamous cell carcinoma cell lines and a novel patient-derived xenograft

doi: 10.1038/s41598-023-35811-3

Figure Lengend Snippet: Eribulin induces cell cycle arrest at the G2/M phase in cSCC cell lines. cSCC cells (A431 and DJM-1) and NHDF were cultured with or without 0.5 nM of eribulin for 24 h. Cells were fixed in 70% ethanol and treated with RNase A. Cells were stained with DNA-binding fluorochrome propidium iodide. DNA contents were determined by fluorescence-activated cell sorting (FACS) analysis. The x-axis indicates propidium iodide fluorescence; the y-axis indicates cell counts. Data are representative of duplicate experiments.

Article Snippet: The human cSCC A431 cell line and normal human primary dermal fibroblasts (NHDFs) were purchased from the American Type Culture Collection (ATCC).

Techniques: Cell Culture, Staining, Binding Assay, Fluorescence, FACS

Journal: Scientific Reports

Article Title: Eribulin inhibits growth of cutaneous squamous cell carcinoma cell lines and a novel patient-derived xenograft

doi: 10.1038/s41598-023-35811-3

Figure Lengend Snippet: Administration of eribulin inhibits tumor growth in vivo. A431 ( A ) and DJM-1 ( B ) cells (5 × 10 6 ) were injected subcutaneously into the bilateral flanks of nu/nu mice. When the tumors reached 8 mm in diameter, eribulin was administrated via the tail vein once a week. Tumor volumes were measured twice per week and were estimated based on the formula: (long axis × short axis 2 )/2. The tumors were extracted and measured at 28 days after treatment initiation. The results are shown as mean ± SD. ** P < 0.01, *** P < 0.001.

Article Snippet: The human cSCC A431 cell line and normal human primary dermal fibroblasts (NHDFs) were purchased from the American Type Culture Collection (ATCC).

Techniques: In Vivo, Injection

Journal: Scientific Reports

Article Title: Eribulin inhibits growth of cutaneous squamous cell carcinoma cell lines and a novel patient-derived xenograft

doi: 10.1038/s41598-023-35811-3

Figure Lengend Snippet: Schematic of the establishment of the cSCC-PDX mouse model. ( A ) The patient’s primary skin lesion was located on the ring finger of the right hand (upper left). Lymph node metastasis was observed 2 years after the first surgery (upper middle). The extracted tumor from the lymph node metastasis was transplanted into NOD/SCID mice (G0, upper right). The enlarged xenografted tumors were transplanted to further generations for primary cell cultures ( B ) and for treatment study ( C ). In HE staining and bright-field images, the cSCC-PDX tissue exhibits similar morphology to that of the patient’s tissue (metastatic lymph node). ( C ) In treatment experiments, tumor-bearing NOD/SCID mice were randomized into a control group, a cisplatin group (5 mg/kg/week), and an eribulin group (1.5 mg/kg/day).

Article Snippet: The human cSCC A431 cell line and normal human primary dermal fibroblasts (NHDFs) were purchased from the American Type Culture Collection (ATCC).

Techniques: Staining, Control

Journal: Scientific Reports

Article Title: Eribulin inhibits growth of cutaneous squamous cell carcinoma cell lines and a novel patient-derived xenograft

doi: 10.1038/s41598-023-35811-3

Figure Lengend Snippet: Cytotoxic agents including eribulin suppress tumor growth in the novel cSCC-PDX mouse model. Tumor-bearing NOD/SCID mice were randomly assigned to a control group (grey line), a cisplatin group (orange line), or an eribulin group (red line). Cisplatin (n = 4, 5 mg/kg) was administered intraperitoneally once per week. Eribulin (n = 4, 1.5 mg/kg) was administered intravenously once per week. ( A ) Tumor volumes were measured twice per week. ( B ) The tumors were extracted and measured at 42 days after treatment initiation. The results are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The human cSCC A431 cell line and normal human primary dermal fibroblasts (NHDFs) were purchased from the American Type Culture Collection (ATCC).

Techniques: Control

Journal: Scientific Reports

Article Title: Eribulin inhibits growth of cutaneous squamous cell carcinoma cell lines and a novel patient-derived xenograft

doi: 10.1038/s41598-023-35811-3

Figure Lengend Snippet: Ki-67 staining indicates that anti-neoplastic agents are effective against the cSCC-PDX tumors. To assess the therapeutic response, treated tumors were extracted at day 42 for all groups (the control, cisplatin, and eribulin groups). ( A ) The panel shows representative images of HE and Ki-67 staining of control and treated SCC-PDX tumors. ( B ) The bar chart indicates the Ki-67 index (the Ki-67-positive cell ratio (%)). The results are shown as mean ± SD. *** P < 0.001.

Article Snippet: The human cSCC A431 cell line and normal human primary dermal fibroblasts (NHDFs) were purchased from the American Type Culture Collection (ATCC).

Techniques: Staining, Clinical Proteomics, Control

Journal: Scientific Reports

Article Title: Eribulin inhibits growth of cutaneous squamous cell carcinoma cell lines and a novel patient-derived xenograft

doi: 10.1038/s41598-023-35811-3

Figure Lengend Snippet: Schematic of the anti-tumor effect of eribulin on the cSCC-PDX. ( A ) In normal keratinocytes, p53 controls the G1/S and G2/M cell cycle checkpoints, and ARID2 regulates the G1/S phase transition. ( B , C ) In the cSCC-PDX, loss of function in p53 and ARID2 leads to the disruption of cell cycle checkpoint controls, which promotes cancer development. Eribulin induces G2/M cell cycle arrest and apoptosis in cSCC cells. The tumors with a loss of tumor suppressor function (such as p53 and ARID2) are particularly sensitive to eribulin.

Article Snippet: The human cSCC A431 cell line and normal human primary dermal fibroblasts (NHDFs) were purchased from the American Type Culture Collection (ATCC).

Techniques: Sublimation, Disruption