607fcf9155358c861d60bf0dc206e3a0 cancers_a2020v12n11p3269.pdf 83a5830b23dbf67eb87eaf6edaf05f416698da05 cancers_a2020v12n11p3269.pdf 85c9d4f85b0a20ea7a29faebeb5aa1ba0826a24efe619048bf7e11ea6ef2e993 cancers_a2020v12n11p3269.pdf Title: TSPAN1: A Novel Protein Involved in Head and Neck Squamous Cell Carcinoma Chemoresistance Subject: Sensitization of resistant cells and cancer stem cells (CSCs) represents a major challenge in cancer therapy. A proteomic study revealed tetraspanin-1 (TSPAN1) as a protein involved in acquisition of cisplatin (CDDP) resistance (Data are available via ProteomeXchange with identifier PXD020159). TSPAN1 was found to increase in CDDP-resistant cells, CSCs and biopsies from head and neck squamous cell carcinoma (HNSCC) patients. TSPAN1 depletion in parental and CDDP-resistant HNSCC cells reduced cell proliferation, induced apoptosis, decreased autophagy, sensitized to chemotherapeutic agents and inhibited several signaling cascades, with phospho-SRC inhibition being a major common target. Moreover, TSPAN1 depletion in vivo decreased the size and proliferation of parental and CDDP-resistant tumors and reduced metastatic spreading. Notably, CDDP-resistant tumors showed epithelial–mesenchymal transition (EMT) features that disappeared upon TSPAN1 inhibition, suggesting a link of TSPAN1 with EMT and metastasis. Immunohistochemical analysis of HNSCC specimens further revealed that TSPAN1 expression was correlated with phospho-SRC (pSRC), and inversely with E-cadherin, thus reinforcing TSPAN1 association with EMT. Overall, TSPAN1 emerges as a novel oncogenic protein and a promising target for HNSCC therapy. Keywords: cancer; HNSCC; resistance; cancer stem cells; apoptosis; autophagy Author: Yoelsis Garcia-Mayea, Cristina Mir, Laia Carballo, Josep Castellvi, Jordi Temprana-Salvador, Juan Lorente, Sergi Benavente, Juana M. García-Pedrero, Eva Allonca, Juan P. Rodrigo and Matilde E. LLeonart Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Thu Nov 12 05:18:46 2020 CET ModDate: Thu Nov 12 06:21:09 2020 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 20 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4734247 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- WLENKM+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 XAWDEN+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 DXVXEO+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 APWWTN+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 OFLXAT+Rpxr Type 1 Builtin yes yes no 28 0 LCJEVJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 37 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 59 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 63 0 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2023-03-31 11:39:37 CEST Size: 4734247 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 490 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Cell Lines and Culture Conditions Destination: subsection.2.1 Item: Title: Drug Assay Destination: subsection.2.2 Item: Title: Proteomic Study Destination: subsection.2.3 Item: Title: Sample Preparation and LC-MS/MS Analyses Destination: subsection.2.4 Item: Title: Peptide 10-Plex TMT Labeling Destination: subsection.2.5 Item: Title: Peptide Fractionation Destination: subsection.2.6 Item: Title: NanoLC-(Orbitrap) MS Analysis Destination: subsection.2.7 Item: Title: Protein Analysis Destination: subsection.2.8 Item: Title: Transfections Destination: subsection.2.9 Item: Title: Colony Formation Assay Destination: subsection.2.10 Item: Title: Quantitative Real-Time PCR (qRT-PCR) Destination: subsection.2.11 Item: Title: Animal Models Destination: subsection.2.12 Item: Title: Image Platform Destination: subsection.2.13 Item: Title: Patients Destination: subsection.2.14 Item: Title: IHC Destination: subsection.2.15 Item: Title: Experimental Design and Statistical Rationale Destination: subsection.2.16 Item: Title: Results Destination: section.3 Children: Item: Title: TSPAN1 Protein Is Upregulated in CDDP-Resistant HNSCC Cells Destination: subsection.3.1 Item: Title: TSPAN1 Inhibition Induces Sensitivity of HNSCC Cells to Chemotherapeutic Agents Destination: subsection.3.2 Item: Title: TSPAN1 Depletion Induces Apoptosis in HNSCC Cells Destination: subsection.3.3 Item: Title: TSPAN1 Depletion Downregulates Several Signaling Cascades with SRC Kinase Signaling as a Central Node Destination: subsection.3.4 Item: Title: TSPAN1 Depletion Decreases Tumor Formation and Metastatic Capacity in Mice Destination: subsection.3.5 Item: Title: TSPAN1 Expression in Patient Samples. Correlations with Active SRC and EMT Features Destination: subsection.3.6 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: TSPAN1: A Novel Protein Involved in Head and Neck Squamous Cell Carcinoma Chemoresistance Author: Yoelsis Garcia-Mayea, Cristina Mir, Laia Carballo, Josep Castellvi, Jordi Temprana-Salvador, Juan Lorente, Sergi Benavente, Juana M. García-Pedrero, Eva Allonca, Juan P. Rodrigo and Matilde E. LLeonart Subject: Sensitization of resistant cells and cancer stem cells (CSCs) represents a major challenge in cancer therapy. A proteomic study revealed tetraspanin-1 (TSPAN1) as a protein involved in acquisition of cisplatin (CDDP) resistance (Data are available via ProteomeXchange with identifier PXD020159). TSPAN1 was found to increase in CDDP-resistant cells, CSCs and biopsies from head and neck squamous cell carcinoma (HNSCC) patients. TSPAN1 depletion in parental and CDDP-resistant HNSCC cells reduced cell proliferation, induced apoptosis, decreased autophagy, sensitized to chemotherapeutic agents and inhibited several signaling cascades, with phospho-SRC inhibition being a major common target. Moreover, TSPAN1 depletion in vivo decreased the size and proliferation of parental and CDDP-resistant tumors and reduced metastatic spreading. Notably, CDDP-resistant tumors showed epithelial mesenchymal transition (EMT) features that disappeared upon TSPAN1 inhibition, suggesting a link of TSPAN1 with EMT and metastasis. Immunohistochemical analysis of HNSCC specimens further revealed that TSPAN1 expression was correlated with phospho-SRC (pSRC), and inversely with E-cadherin, thus reinforcing TSPAN1 association with EMT. Overall, TSPAN1 emerges as a novel oncogenic protein and a promising target for HNSCC therapy. 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