Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: Association between the clinicopathological variables and BTC expression in 38 OSCC patients.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Expressing, Virus, Infection

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: DEGs in OSCC. (A,B) Volcano plots were constructed using FC values and FDRs. The red points in the plot represent the overexpressed mRNAs, and the blue points indicate the downregulated mRNAs with statistical significance. (A) DEGs between normal and tumor tissues. (B) DEGs between metastasis-positive and metastasis-negative tumor tissues. (C) Hierarchical clustering analysis of mRNAs that were differentially expressed between metastasis-negative and metastasis-positive tissues. The normalized expression levels in the heatmaps are colored from blue to red in ascending order. (D) Multivariate Cox regression analysis according to gene expression. (E) Kaplan–Meier survival curves were performed to show the prognosis of patients with high and low expression of BTC through the analysis of the mRNA expression profile data of 260 OSCC tumor samples from TCGA database.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Construct, Expressing, Gene Expression

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: BTC expression is correlated with clinicopathological parameters in OSCC patients. (A) Representative images of immunohistochemical staining and (B) immunoreactive score of BTC in human normal mucosa samples, OSCC tissue samples, and metastatic LN samples. The experiment was repeated three times independently. Results are shown as mean ± SD. T-test, n = 38. (C) mRNA expression of BTC in normal and tumor tissues was detected by RT-PCR. (D) Kaplan–Meier survival curves of 38 patients from our department. (E–L) Analysis of 330 OSCC samples from TCGA database and 32 pairs of OSCC samples selected from TCGA database showed the comparison of (E,F) BTC expressed in tumor tissues and normal tissues. Correlation analysis with BTC expression and (G) age, (H) histological grade, (I) sex, (J) T category, (K) tumor stage, and (L) LN metastasis status. * p < 0.05. ** p < 0.01. *** p < 0.001. **** p < 0.0001.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Expressing, Immunohistochemical staining, Staining, Reverse Transcription Polymerase Chain Reaction, Comparison

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: Statistical analyses of clinicopathological features associated with survival in 38 OSCC patients with the multivariate Cox proportional hazards models.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Expressing

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: Overexpression of BTC inhibits the proliferation, migration, and invasion of OSCC cell lines. (A) Cancer cell transfectants of the BTC-expressing vector and empty vector control were identified in SCC4 and CAL27 cells by Western blot. (B,C) Overexpression of BTC inhibited cell proliferation, as indicated by the CCK-8 assay, in CAL27 and SCC4 cells. (D) Wound healing assay showed that overexpression of BTC inhibited CAL27 and SCC4 cell migration. (E,F) Transwell assays showed that the migration and invasion abilities of CAL27 and SCC4 cells were impaired after overexpression of BTC.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Over Expression, Migration, Expressing, Plasmid Preparation, Control, Western Blot, CCK-8 Assay, Wound Healing Assay

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: (A) Comparison of EMT scores in normal and tumor (in BTC low- and high-expression groups) tissue. (B) TCGA and GSEA showed highly regulated genes in patients with high-BTC expression versus those with low-BTC expression. (C) Heatmap of EMT marker expression in the BTC high- and low-expression groups. (D–F) Relationship between BTC and EMT markers, such as E-cadherin, N-cadherin, and vimentin. (G) PPI network analysis and Western blot analysis. The PPI network of the DEGs was constructed using STRING. The network nodes represent different proteins. The edges represent protein–protein associations, and the line thickness indicates the strength of the supporting data. (H) Protein expression level of EMT-related markers and the PI3K-AKT signaling pathway after overexpression of BTC in SCC4 and Cal27 cells.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Comparison, Expressing, Marker, Western Blot, Construct, Over Expression

Journal: eLife

Article Title: Reprogramming and redifferentiation of mucosal-associated invariant T cells reveal tumor inhibitory activity

doi: 10.7554/eLife.70848

Figure Lengend Snippet: ( A ) Time course of 5-(2-oxopropylideneamino)-6-D-ribitylaminouracil (5-OP-RU)-dependent MR1 expression. The indicated cancer cell lines were challenged with 5-OP-RU. MR1 expression levels on the cell surface at the indicated time point are shown as relative geometric mean fluorescent intensity (gMFI). Data are representative of three independent experiments. ( B ) m-reMAIT cell dose-dependent survival extension. C57BL/6 mice received the indicated amounts of m-reMAIT cells 6 days prior to the Lewis lung carcinoma (LLC) inoculation (3 × 10 5 cells/mouse i.v.), and survival was monitored (n = 10–12/group). Data are representative of three independent experiments. p-Values between the indicated group are shown (the log-rank test). ( C ) Effects of the multiple transfers of m-reMAIT cells on survival. The survival of C57BL/6 mice that received m-reMAIT cells (1 × 10 6 /mouse, i.p.) 6 days prior to the LLC inoculation (3 × 10 5 cells/mouse, i.v.), and of mice that received LLC and two more consecutive transfers of m-reMAIT cells (1 × 10 6 /transfer/mouse) was monitored (n = 10–12/group). Sham-treated mice that only received LLC served as a control. Data are representative of two independent experiments. p-Values between the indicated groups are shown (the log-rank test). ( D ) Effects of m-reMAIT cells on in situ tumor growth. Growth curve of LLC. LLC (3 × 10 5 /mouse) was subcutaneously inoculated into the right flank of C57BL/6 mice 6 days after the m-reMAIT cell transfer (i.p.). Tumor size was plotted with time. Sham treated (●), 0.3 × 10 6 transferred (■), 1.0 × 10 6 transferred (▲), and 3.0 × 10 6 m-reMAIT cells transferred (▼). Data are shown as SEM (5–6 mice per group). Figure 3—source data 1. Time-dependent MR1 expression in various cancer cell lines upon 5-(2-oxopropylideneamino)-6-D-ribitylaminouracil (5-OP-RU) challenge. ( A ), m-reMAIT cell dose-dependent mouse survival ( B ). Effects of the multiple transfers of m-reMAIT cells on mouse survival ( C ). Effects of m-reMAIT cell dose on in situ tumor growth ( D ).

Article Snippet: Cell line ( M. musculus ) , Lewis lung carcinoma (LLC) , Riken Bioresource Center , Cat# RCB0558; RRID :CVCL_4358 , .

Techniques: Expressing, Control, In Situ

Journal: eLife

Article Title: Reprogramming and redifferentiation of mucosal-associated invariant T cells reveal tumor inhibitory activity

doi: 10.7554/eLife.70848

Figure Lengend Snippet: The number of m-reMAIT cells from the indicated tissues upon adoptive transfer in the recipient is shown. Tissue samples were prepared at the indicated time point with or without subcutaneous Lewis lung carcinoma (LLC) injection (n = 3–4). MAIT: mucosal-associated invariant T cell. Figure 3—figure supplement 1—source data 1. Delayed emergence of m-reMAIT cells in the skin upon adoptive transfer.

Article Snippet: Cell line ( M. musculus ) , Lewis lung carcinoma (LLC) , Riken Bioresource Center , Cat# RCB0558; RRID :CVCL_4358 , .

Techniques: Adoptive Transfer Assay, Injection

Journal: eLife

Article Title: Reprogramming and redifferentiation of mucosal-associated invariant T cells reveal tumor inhibitory activity

doi: 10.7554/eLife.70848

Figure Lengend Snippet: ( A ) Activation of m-reMAIT cells by NK cells. CD69 expression in m-reMAIT cells (reMAIT ●) and NK cells (NK ◯) upon incubation at the indicated ratio. The percentage of cells expressing CD69 (left panel) and the intensity of CD69 in each cell population (right panel) are shown. ( B ) Cytokines and chemokines upon a coculture. Cytokines and chemokines released upon a coculture of m-reMAIT cells and NK cells at the indicated ratio are shown (reMAIT:NK). Amounts were quantified with LegendPlex. Representative data from two independent experiments are shown. ( C ) Transcripts relevant to cytolytic activity in m-reMAIT cells. Ifng , Gzma , Gzmb , Tbf , Gzmk , Pfr1 , Fasl , Tnfsf10 , Il6 , Il17a , Il22 , Ccl3 , Ccl4 , Ccl5 , and Ccl22 in m-reMAIT cells cultured with NK cells were quantified with qRT-PCR. m-reMAIT cells and NK cells were sort-purified after the coculture (purity >98%) or cultured individually. The expression of each transcript was normalized with Gapdh, and fold changes in the relative expression of the transcript in m-reMAIT cells cultured with NK cells relative to that in m-reMAIT cells cultured alone are shown. Data are representative of three independent experiments. ( D ) Transcripts relevant to cytolytic activity in NK cells. Fold changes in the relative expression of the indicated transcript as described in ( C ) in NK cells cultured with m-reMAIT cells relative to that in NK cells alone are shown. Representative data from three independent experiments are shown. ( E ) Activation and degranulation of NK cells and m-reMAIT cells. The expression of CD69, an activation marker, and CD107a, a marker for the exocytosis of cytolytic granules, was assessed under various culture conditions. The percentages of CD69 + cells and CD69 + CD107a + cells among NK cells alone (control), NK cells cocultured with m-reMAIT cells (+reMAIT), NK cells cultured with Yac-1 (+Yac-1), and NK cells cocultured with m-reMAIT cells and Yac-1 (+reMAIT/Yac-1) (upper panels). The percentages of CD69 + cells and CD69 + CD107a + cells among m-reMAIT cells alone (control), m-reMAIT cells cocultured with NK cells (+NK), m-reMAIT cells cocultured with Yac-1 (+Yac-1), and m-reMAIT cells cocultured with NK cells and Yac-1 (+NK/Yac-1) (lower panels). Data are representative of three independent experiments. ( F ) Cytolytic activity against Yac-1. Cytolytic activity of m-reMAIT cells (reMAIT ◯), NK cells (NK □), and NK cells plus m-reMAIT cells (NK+reMAIT ●). Cytolytic activities (% lysis) at different effector (NK cells, m-reMAIT cells, and NK cell+m-reMAIT cells)/Target (Yac-1) (E/T) ratios are shown. Representative data from three experiments are shown. The significance of differences between the groups at the indicated E/T ratio assessed with a two-way ANOVA is shown (*p<0.05, **p<0.01, ***p<0.005). From the top, NK+reMAIT vs. reMAIT, NK+reMAIT vs. NK, and reMAIT vs. NK. Data are representative of three independent experiments. ( G ) Cytolytic activity against Lewis lung carcinoma (LLC). The cytolytic activities of m-reMAIT cells (◯), NK cells (□), and m-reMAIT cells plus NK cells (●) against LLC at the indicated E/T ratio are shown as % lysis. The significance of differences between the groups is calculated as in ( E ). Data are representative of three independent experiments. ( H ) NK cell-dependent extension of survival. C57BL/6 mice were divided into two groups, one that received 1 × 10 6 m-reMAIT cells (reMAIT) and another that was left untreated (n). Each group was further divided into two subgroups, one that received consecutive injections of anti-Asialo GM1 (AsG) (–1 and 16 days, 50 μg/mouse) after the LLC inoculation (3 × 10 5 i.v.) and another that was left untreated (n). Survival was monitored thereafter. Representative data from two independent experiments are shown (n = 10–14/group). p-Values between the indicated groups are shown (the log-rank test). Figure 4—source data 1. Antitumor activity of m-reMAIT cells bolstered by NK cells. Activation of m-reMAIT cells by NK cells ( A ). Cytokines and chemokines produced upon a coculture ( B ). Transcripts relevant to cytolytic activity in m-reMAIT cells ( C ). Transcripts relevant to cytolytic activity in NK cells ( D ). Activation and degranulation of NK cells and m-reMAIT cells ( E ). Cytolytic activity against Yac-1 ( F ). Cytolytic activity against LLC ( G ). NK cell-dependent extension of survival ( H ).

Article Snippet: Cell line ( M. musculus ) , Lewis lung carcinoma (LLC) , Riken Bioresource Center , Cat# RCB0558; RRID :CVCL_4358 , .

Techniques: Activation Assay, Expressing, Incubation, Activity Assay, Cell Culture, Quantitative RT-PCR, Purification, Marker, Control, Lysis, Produced

Journal: eLife

Article Title: Reprogramming and redifferentiation of mucosal-associated invariant T cells reveal tumor inhibitory activity

doi: 10.7554/eLife.70848

Figure Lengend Snippet:

Article Snippet: Cell line ( M. musculus ) , Lewis lung carcinoma (LLC) , Riken Bioresource Center , Cat# RCB0558; RRID :CVCL_4358 , .

Techniques: Virus, Flow Cytometry, Purification, Blocking Assay, Control, Western Blot, In Vivo, Sequencing, Southern Blot, Recombinant, Cell Stimulation, Cell Isolation, Software