Journal: Cancers

Article Title: WISP-1 Promotes Epithelial-Mesenchymal Transition in Oral Squamous Cell Carcinoma Cells via the miR-153-3p/Snail Axis

doi: 10.3390/cancers11121903

Figure Lengend Snippet: WISP-1 induces epithelial-to-mesenchymal transition (EMT) functioning via E-cadherin downregulation. ( A , B ) SCC4 cells were treated with different concentrations of WISP-1 (0–30 ng/mL) for 24 h; cell migration and cell-scattering phenotype were analyzed by wound healing and scatter assays, respectively. Images of living cells were captured by microscope and the width of the gap in each scratch was measured by ImageJ software. ( C , D ) Cells were incubated with different concentrations of WISP-1 (0–30 ng/mL) for 24 h and EMT marker expression was evaluated by qPCR and Western blot assay. The α-tubulin content was used to normalize for levels of E-cadherin protein. ( E ) E-cadherin mRNA expression in tumor tissue and adjacent normal tissue was determined in TCGA database records. Mean levels (ranges) of mRNA expression in normal and tumor samples, respectively: 10,660 (2858.6–17,763.1), 9095 (4.929–18,812.1); log 2 (fold-change): –0.229. Results are expressed as the mean ± SEM. * p < 0.05 compared with the control group.

Article Snippet: The human OSCC cell line SCC4 was purchased from the Bioresource Collection and Research Center (BCRC; Hsinchu, Taiwan).

Techniques: Migration, Microscopy, Software, Incubation, Marker, Expressing, Western Blot, Control

Journal: Cancers

Article Title: WISP-1 Promotes Epithelial-Mesenchymal Transition in Oral Squamous Cell Carcinoma Cells via the miR-153-3p/Snail Axis

doi: 10.3390/cancers11121903

Figure Lengend Snippet: WISP-1 stimulates the EMT process via the integrin αvβ3 receptor in oral squamous cell carcinoma (OSCC) cells. ( A , B ) SCC4 cells were pretreated with Arg–Gly–Asp (RGD) (100 nM) or Arg–Ala–Asp (RAD) (100 nM) for 30 min, then stimulated with WISP-1 for 24 h. E-cadherin expression was examined by IF and qPCR assays. ( C ) Cells were pretreated with RGD or RAD for 30 min then stimulated with WISP-1 for 24 h, and cell migration was examined by the wound healing assay. Results are expressed as the mean ± SEM. * p < 0.05 compared with controls; # p < 0.05 compared with the WISP-1-treated group.

Article Snippet: The human OSCC cell line SCC4 was purchased from the Bioresource Collection and Research Center (BCRC; Hsinchu, Taiwan).

Techniques: Expressing, Migration, Wound Healing Assay

Journal: Cancers

Article Title: WISP-1 Promotes Epithelial-Mesenchymal Transition in Oral Squamous Cell Carcinoma Cells via the miR-153-3p/Snail Axis

doi: 10.3390/cancers11121903

Figure Lengend Snippet: Involvement of FAK and ILK in WISP-1-regulated EMT functioning. ( A , B ) SCC4 cells were incubated with WISP-1 (30 ng/mL) for the indicated time intervals; FAK and ILK activation was examined by Western blot assay. ( C , D ) Cells were pretreated with RGD (100 nM) for 30 min then treated with WISP-1 for 10 min; FAK and ILK activation was examined by Western blot assay. FAK and GSK3β content was used to normalize for pFAK and pGSK3βlevels. ( E , F ) Cells were pretreated for 30 min with a FAKi (10 M) and KP392 (10 M), prior to incubation with WISP-1 for 24 h. E-cadherin expression was examined by IF and qPCR assays. ( G ) Cells were pretreated for 30 min with a FAKi (10 M) and KP392 (10 M), then incubated with WISP-1 for 24 h. Cell migration was examined by the wound healing assay. Results are expressed as the mean ± SEM. * p < 0.05 compared with controls; # p < 0.05 compared with the WISP-1-treated group.

Article Snippet: The human OSCC cell line SCC4 was purchased from the Bioresource Collection and Research Center (BCRC; Hsinchu, Taiwan).

Techniques: Incubation, Activation Assay, Western Blot, Expressing, Migration, Wound Healing Assay

Journal: Cancers

Article Title: WISP-1 Promotes Epithelial-Mesenchymal Transition in Oral Squamous Cell Carcinoma Cells via the miR-153-3p/Snail Axis

doi: 10.3390/cancers11121903

Figure Lengend Snippet: WISP-1 induces Akt phosphorylation via the integrin αvβ3/FAK/ILK signaling pathway. ( A ) SCC4 cells were incubated with WISP-1 (30 ng/mL) for the indicated time intervals; levels of Akt phosphorylation were examined by Western blot assay. ( B , C ) Cells were pretreated with RGD (100 nM), a FAKi (10 M), or KP392 (10 M) for 30 min, then incubated with WISP-1 for 30 min. Akt activation was examined by Western blot assay. Akt protein was used to normalize for pAkt levels. ( D – F ) Cells were pretreated for 30 min with Akti (10 M), prior to incubation with WISP-1 for 24 h. E-cadherin expression was examined by IF and qPCR assays. Cell migration was examined by the wound healing assay. Results are expressed as the mean ± SEM. * p < 0.05 compared with the control group; # p < 0.05 compared with the WISP-1-treated group.

Article Snippet: The human OSCC cell line SCC4 was purchased from the Bioresource Collection and Research Center (BCRC; Hsinchu, Taiwan).

Techniques: Phospho-proteomics, Incubation, Western Blot, Activation Assay, Expressing, Migration, Wound Healing Assay, Control

Journal: Cancers

Article Title: WISP-1 Promotes Epithelial-Mesenchymal Transition in Oral Squamous Cell Carcinoma Cells via the miR-153-3p/Snail Axis

doi: 10.3390/cancers11121903

Figure Lengend Snippet: WISP-1 induces EMT function by Snail up-regulation in OSCC. ( A , B ) Cells were incubated with different concentrations of WISP-1 (0–30 ng/mL) for 24 h, then Snail and Twist expression were examined by qPCR and Western blot assays. The α-tubulin protein was used to normalize for levels of Snail and Twist. ( C , D ) Cells were pretreated for 30 min with RGD, FAKi, KP392, or Akti, prior to incubation with WISP-1 for 24 h. Snail translocation into the nucleus was examined by IF assay. Snail mRNA expression was examined by qPCR assay. ( E ) Cells were transfected with Snail siRNA (10 μM) or control siRNA (10 μM) for 24 h then stimulated with WISP-1 for 24 h; E-cadherin mRNA expression was examined using the qPCR assay. ( F ) Snail mRNA expression in tumor tissue and adjacent normal tissue was analyzed using records from the TCGA database. Mean levels (ranges) of Snail mRNA expression in normal and tumor samples, respectively: 6.064 (2.53–8.719), 6.555 (2.594–9.079); log 2 (fold-change): 0.491. Results are expressed as the mean ± SEM. * p < 0.05 compared with controls; # p < 0.05 compared with the WISP-1-treated group.

Article Snippet: The human OSCC cell line SCC4 was purchased from the Bioresource Collection and Research Center (BCRC; Hsinchu, Taiwan).

Techniques: Incubation, Expressing, Western Blot, Translocation Assay, Transfection, Control

Journal: Cancers

Article Title: WISP-1 Promotes Epithelial-Mesenchymal Transition in Oral Squamous Cell Carcinoma Cells via the miR-153-3p/Snail Axis

doi: 10.3390/cancers11121903

Figure Lengend Snippet: A schematic model depicting how WISP-1 regulates EMT functioning in OSCC cells. The modeling shows two potential signaling pathways underlying OSCC-derived WISP-1 regulation of EMT activation: (1) WISP-1 induces Snail expression via the integrin αvβ3/FAK/ILK/Akt signaling pathway, which in turn activates EMT functioning; (2) WISP-1 also inhibits miR-153-3p-induced downregulation of Snail mRNA translation and thus stimulates Snail protein expression and EMT activation. (RGD peptide: a target epitope of integrin αvβ3)

Article Snippet: The human OSCC cell line SCC4 was purchased from the Bioresource Collection and Research Center (BCRC; Hsinchu, Taiwan).

Techniques: Protein-Protein interactions, Derivative Assay, Activation Assay, Expressing

Journal: JCI Insight

Article Title: CMTM6 drives cisplatin resistance by regulating Wnt signaling through the ENO-1/AKT/GSK3 β axis

doi: 10.1172/jci.insight.143643

Figure Lengend Snippet: ( A ) Schematic representation of sensitive, early, and late cisplatin-resistant OSCC line for global proteomic profiling. The establishment of sensitive, early, and late resistant cells is described in Methods. ( B ) The lysates were isolated from parental sensitive (H357CisS), early (H357CisR4M), and late (H357CisR8M) cisplatin-resistant cells and subjected to global proteomic profiling. The schematic diagram depicts the iTRAQ labeling strategy for proteomic analysis. 0R11 and 0R12 are biological replicates of the H357CisS group, 4R11: 4R12 and 4R2 are technical and biological replicates of the H357CisR4M group, and 8R11: 8R12 and 8R2 are technical and biological replicates of H357CisR8M group. ( C ) Principal component analysis (PCA) of global proteomic profiling sensitive, early (4M), and late resistant cells (8M). ( D ) Volcano plot indicating deregulated genes in proteome profiling of sensitive and cisplatin-resistant cells. CMTM6 is the top-ranked upregulated genes in 4M and 8M cisplatin-resistant groups. SCX, strong cation exchange; and LC-ESI, liquid chromatography electrospray ionization.

Article Snippet: H357, SCC9, and SCC4 (human tongue OSCC) cell lines were obtained from MilliporeSigma, sourced from a European collection of authenticated cell culture.

Techniques: Isolation, Multiplex sample analysis, Labeling, Liquid Chromatography

Journal: JCI Insight

Article Title: CMTM6 drives cisplatin resistance by regulating Wnt signaling through the ENO-1/AKT/GSK3 β axis

doi: 10.1172/jci.insight.143643

Figure Lengend Snippet: ( A ) Cell lysates from indicated resistant and sensitive OSCC cells were isolated and subjected to immunoblotting ( n = 3) against CMTM6 and β-actin antibodies. ( B ) Relative mRNA (fold change) CMTM6 expression was analyzed by quantitative real-time PCR (qRT-PCR) in indicated cells (mean ± SEM, n = 3), * P < 0.05 by 1-way ANOVA. ( C ) Cell lysates from indicated resistant and sensitive OSCC cells were isolated and subjected to immunoblotting ( n = 3) against CMTM6 and β-actin antibodies. ( D ) Relative mRNA (fold change) CMTM6 expression was analyzed by qRT-PCR in indicated cells (mean ± SEM, n = 3), * P < 0.05 by 1-way ANOVA. ( E ) Relative mRNA expression of CMTM6 was analyzed by qRT-PCR in different chemotherapy nonresponder (CT nonresponder) OSCC tumors as compared with CT responder tumors (median, n = 11 for CT responder and n = 23 for CT nonresponder). * P < 0.05 by 2-tailed Student’s t test. ( F ) Protein expression of CMTM6 was analyzed by IHC in CT responder and CT nonresponder OSCC tumors. Scale bars: 50 μm. ( G ) IHC scoring for CMTM6 from J (Q score = staining intensity × percent of staining) (median, n = 11 for CT responder and n = 23 for CT nonresponder). * P < 0.05 by 2-tailed Student’s t test. ( H ) CMTM6 protein expression was analyzed by IHC in pre– and post–TPF-treated paired tumor samples for CT nonresponder patients. Scale bars: 50 μm. ( I ) IHC scoring for CMTM6 from H (Q score = staining intensity × percent of IHC staining). ( J ) Relapse-free survival (RFS) plot for CMTM6 using Kaplain-Meier Plotter.

Article Snippet: H357, SCC9, and SCC4 (human tongue OSCC) cell lines were obtained from MilliporeSigma, sourced from a European collection of authenticated cell culture.

Techniques: Isolation, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Staining, Immunohistochemistry

Journal: JCI Insight

Article Title: CMTM6 drives cisplatin resistance by regulating Wnt signaling through the ENO-1/AKT/GSK3 β axis

doi: 10.1172/jci.insight.143643

Figure Lengend Snippet: ( A ) Cisplatin-resistant OSCC lines were stably transfected with NTShRNA and CMTM6ShRNA as described in Methods. ShRNA#1 targets CMTM6 mRNA and ShRNA#2 targets 5′UTR of CMTM6 mRNA. Lysates were collected from indicated stable clone, and immunoblotting ( n = 3) was performed with anti-CMTM6 and β-actin antibodies. ( B ) Cisplatin-resistant cells stably expressing NTShRNA and CMTM6ShRNA were treated with cisplatin for 12 days and colony-forming assays performed as described in Methods (left). Relative colony number ( n = 3, * P < 0.05 by 2-way ANOVA). Photographs of colony-forming assay in each group (right). ( C ) Cisplatin-resistant cells stably expressing NTShRNA and CMTM6ShRNA were treated with cisplatin for 48 hours, and cell viability was determined by MTT assay ( n = 3), * P < 0.05 by 2-way ANOVA.

Article Snippet: H357, SCC9, and SCC4 (human tongue OSCC) cell lines were obtained from MilliporeSigma, sourced from a European collection of authenticated cell culture.

Techniques: Stable Transfection, Transfection, shRNA, Western Blot, Expressing, MTT Assay

Journal: International Journal of Nanomedicine

Article Title: Future trends and emerging issues for nanodelivery systems in oral and oropharyngeal cancer

doi: 10.2147/IJN.S133219

Figure Lengend Snippet: Use of nanomaterial-based drug-delivery systems in oral cancer

Article Snippet: , Lipid–calcium–phosphate NPs with siVEGFA , Human OSCC, SCC4, and SAS xenograft , .

Techniques: In Vitro, Polymer, shRNA, Transfection, In Vivo, Liposomes