human bladder epithelial cell line 5637  (ATCC)

Journal: BMC Cancer

Article Title: Conjoining cell reprogramming and mass spectrometry to identify the proteomic variations in the reprogrammed bladder cancer cells: finding cues of normalisation

doi: 10.1186/s12885-026-15634-x

Figure Lengend Snippet: Reprogramming process of the bladder cancer cell line HTB-5. A Multiplicity of Infection (MOI) optimisation for Sendai virus was performed using EmGFP Sendai Fluorescence Reporter with ranging MOI concentrations from 3 to 12 and the cultures were monitored for 168 h. Mean fluorescence intensity was calculated using Image J and the change in intensities were shown as bar graphs. HTB-5 cells were compatible with all MOIs but MOI 6 was selected for further experiments. B A visual depiction of reprogramming procedure to create a distinct cell line from the parental bladder cancer cells. Two alternative routes the cells may choose to follow are indicated as partial and full reprogramming. Created in BioRender. Iskender Izgi, B. (2025) https://BioRender.com/qb3h9ze. C HTB-5 cells transfected with SeV-OSKM were monitored and photographed during the reprogramming stages. After transduction, HTB-5 were shown to lose its elongated, multi-sided structure and gained an epithelial-like morphology while forming colonies resembling iPSCs. D At passage 7, the reprogrammed HTB-5 cells were characterised for pluripotency-associated marker expression. The pluripotency markers OCT4, LIN28, SOX-2, c-MYC and NANOG were positively expressed in the reprogrammed HTB-5 cells and newly established cell line was referred as HTB-5 PR. E The parental HTB-5 cells failed to express pluripotency-associated markers. Scale bars represent 100 μm

Article Snippet: Human grade 4 bladder cancer cell line HTB-5 (ATCC TCCSUP, RRID: CVCL_1738) and immortalised uroepithelial cell line SV-HUC-1 (ATCC CRL-9520, RRID: CVCL_3798 ) were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Infection, Virus, Fluorescence, Transfection, Transduction, Marker, Expressing

Journal: BMC Cancer

Article Title: Conjoining cell reprogramming and mass spectrometry to identify the proteomic variations in the reprogrammed bladder cancer cells: finding cues of normalisation

doi: 10.1186/s12885-026-15634-x

Figure Lengend Snippet: Behavioural assays with bladder cancer cells and reprogrammed bladder cancer cells. A Crystal violet staining of colonies of HTB-5 and HTB-5 PR. HTB-5 cells formed smaller and more colonies, while HTB-5 PR cells form fewer but larger colonies (± SD, n = 3, 2-tailed unpaired t-test, HTB-5 v HTB-5 PR, * P < 0.05) B Cell proliferation was assessed using the CCK-8 assay. HTB-5 and HTB-5 PR cells were treated with increasing doses of paclitaxel or with doxorubicin for 36 h. Paclitaxel inhibited the growth of both HTB-5 and HTB-5 PR cells in a dose-dependent manner (upper graph). Doxorubicin inhibited the growth of both HTB-5 and rep HTB-5 in a dose-dependent manner (lower graph). Data are presented as the means ± SD (error bars) from two independent experiments. (Mean ± SD, n = 3, 2-tailed unpaired t-test, HTB-5 v control; ** p < 0.05, * p < 0.01. HTB-5 PR vs. control; ## p < 0.05, # p < 0.01. CCK-8: Cell-Counting Kit-8; Pac: Paclitaxel, Dox: Doxorubicin; SD: standard deviation.) C A clonogenic assay was performed after exposing the cells to increasing doses of doxorubicin or paclitaxel. Colony forming ability was depicted as relative colony number at 560 nm (right). Mean ± SD, n = 3, 2-tailed unpaired t-test, HTB-5 v control; ** p < 0.05, * p < 0.01. HTB-5 PR vs. control; ## p < 0.05, # p < 0.01) D Effect of cancer cell reprogramming on transwell migration and invasion. The number of migrating HTB-5 PR cells was determined by quantifying the amount of crystal violet at an OD at 560 nm (± SD, n = 3, 2-tailed unpaired t-test, HTB-5 v HTB-5 PR, * p < 0.05) E Comparison of migration abilities by wound healing assay. HTB-5 and HTB-5 PR cells were treated with increasing doses of paclitaxel, and cell migration was observed at 24, 48 and 72 h. HTB-5 PR cells migrated to close the wound, while different doses of paclitaxel inhibited cell migration, increased cell death as observed in parental HTB-5 cells. The wound widths are depicted as bar graphs (lower panel)

Article Snippet: Human grade 4 bladder cancer cell line HTB-5 (ATCC TCCSUP, RRID: CVCL_1738) and immortalised uroepithelial cell line SV-HUC-1 (ATCC CRL-9520, RRID: CVCL_3798 ) were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Staining, CCK-8 Assay, Control, Cell Counting, Standard Deviation, Clonogenic Assay, Migration, Comparison, Wound Healing Assay

Journal: BMC Cancer

Article Title: Conjoining cell reprogramming and mass spectrometry to identify the proteomic variations in the reprogrammed bladder cancer cells: finding cues of normalisation

doi: 10.1186/s12885-026-15634-x

Figure Lengend Snippet: Comparative proteomic analysis of bladder cancer cell line HTB-5, reprogrammed bladder cancer cell line HTB-5-PR and human normal bladder epithelial cell line SV-HUC-1. A In total, 3654 proteins were identified with high confidence (false discovery rate < 1%) with at least two or more unique peptide fragments. Data analysis was enhanced by filtering the identified proteins to 1969 proteins that contain at least 5 or more unique peptides. In the hierarchical clustering of proteins with 2-fold change in abundance (FC > 2 or < 0.5), HTB-5 PR cells exhibit a transition state between normal epithelial cells and grade 4 bladder cancer cells. B Heat maps representing the regulated proteins involved in pluripotency, stem cell differentiation and extracellular matrix in each sample. The dendrogram depicts a separation of HTB-5-PR from HTB-5 and SV-HUC-1 cells in terms of pluripotency-associated protein expression. The cluster analysis was performed using Pearson correlation for the distance measure, protein intensities are shown using a colour scale ranging from red (high) to blue (low). The dendrogram above the heatmap shows the degree of relationship between protein profiles in the three samples. Red colour corresponds to higher z-scores. A z-score equivalent to 0 represents the mean abundance value for that protein. C During the reprogramming process, the abundance ratios of HTB-5/SV-HUC-1 were analysed against the expression levels of proteins in the normal bladder epithelial cell line. In the transition phase from grade 4 bladder cancer cell line toward reprogrammed HTB-5 PR cells, a variety of biological processes were shown to be regulated by g: Profiler analysis. D Out of 348 proteins were upregulated and out of 281 proteins downregulated, the expression levels of 295 proteins, which comprise 47% of the regulated proteins, were normalised in HTB-5 cells upon reprogramming

Article Snippet: Human grade 4 bladder cancer cell line HTB-5 (ATCC TCCSUP, RRID: CVCL_1738) and immortalised uroepithelial cell line SV-HUC-1 (ATCC CRL-9520, RRID: CVCL_3798 ) were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Cell Differentiation, Expressing

Journal: BMC Cancer

Article Title: Conjoining cell reprogramming and mass spectrometry to identify the proteomic variations in the reprogrammed bladder cancer cells: finding cues of normalisation

doi: 10.1186/s12885-026-15634-x

Figure Lengend Snippet: STRING network analysis of upregulated and downregulated proteins indicate progress toward normalisation in parental HTB-5 cells. Results of annotated keywords (by UniProt) obtained using functional enrichment analysis in STRING 12.5 (FDR < 0.05), indicating enriched categories and number of proteins mapped. After reprogramming, 47% of total proteins in parental HTB-5 cells that approximate the expression levels to those of the normal bladder epithelial cell line SV-HUC-1 cells were considered to be normalised and analysed for protein-protein interaction. A Protein-protein interactions of upregulated 149 proteins and downregulated 148 proteins (upper panel) and the proteins involved in the GO annotations showing the most clustering are presented at the bottom of each protein-protein interaction network (lower panel). Nodes represent proteins, lines between nodes represent edges showing interactions with a minimum confidence score of 0.7. Each GO annotation is indicated with a different colour, and the nodes are coloured depending on their involvement in certain biological processes. B g: Profiler analysis of upregulated and downregulated proteins during the reprogramming process. The graphs show the enriched terms, most of which are also highlighted in the STRING analysis. The ten most highly upregulated terms (lower left panel) and downregulated terms (lower right panel) are ranked by decreasing protein number detected in each term

Article Snippet: Human grade 4 bladder cancer cell line HTB-5 (ATCC TCCSUP, RRID: CVCL_1738) and immortalised uroepithelial cell line SV-HUC-1 (ATCC CRL-9520, RRID: CVCL_3798 ) were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Functional Assay, Expressing, Protein-Protein interactions

Journal: BMC Cancer

Article Title: Conjoining cell reprogramming and mass spectrometry to identify the proteomic variations in the reprogrammed bladder cancer cells: finding cues of normalisation

doi: 10.1186/s12885-026-15634-x

Figure Lengend Snippet: Functional enrichment analysis of reprogrammed HTB-5 PR cells. A The STRING database was used to construct the protein-protein interaction (PPI) networks of upregulated 145 proteins (left panel) and downregulated 85 proteins (right panel) in HTB-5 PR cells. Nodes represent proteins, and the lines between them depict interactions with a minimum confidence score of 0.7. B g: Profiler analysis of upregulated and downregulated proteins in the reprogrammed HTB-5 PR cells. The graph represents the GO enrichment analysis of upregulated proteins (left panel in shaded red) in the biological process and downregulated proteins (right panel in shaded blue) in the biological process. Ten highly upregulated and downregulated GO terms are ranked by the number of proteins detected in each term, listed in decreasing order. Spontaneous differentiation upon reprogramming leads to an increase in nervous system development markers and to a decrease in epithelium development markers (marked in blue) as detected by both STRING and g: Profiler analysis. C The clustering of differentially expressed proteins involved in neural development and epithelial cell differentiation is displayed in heatmaps. The parental HTB-5 cells and normal epithelial cell line SV-HUC-1 are grouped based on similar protein expression profiles showing downregulation of neuronal markers and upregulation of epithelial markers, while the reprogrammed bladder cancer cell line HTB-5 PR exhibits an opposite expression profile. The data is consistent with the morphological evidence documented during the reprogramming process (from right to left), showing the parental HTB-5 cells transform into pluripotent-like colonies and gain neuron-like trajectories as a result of spontaneous differentiation in time. The protein intensities are shown using a colour scale ranging from red (high) to blue (low). Red colour corresponds to higher z-scores. A z-score equivalent to 0 represents the mean abundance value for that protein

Article Snippet: Human grade 4 bladder cancer cell line HTB-5 (ATCC TCCSUP, RRID: CVCL_1738) and immortalised uroepithelial cell line SV-HUC-1 (ATCC CRL-9520, RRID: CVCL_3798 ) were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Functional Assay, Construct, Cell Differentiation, Expressing