Journal: Oncotarget

Article Title: The inhibition of malignant melanoma cell invasion of bone by the TLR7 agonist R848 is dependent upon pro-inflammatory cytokines produced by bone marrow macrophages

doi: 10.18632/oncotarget.25711

Figure Lengend Snippet: MTS assays of cultures of B16F10 cells, the mammary tumor cell line MMT060562 (MMT), and the lung carcinoma cell line Ex-3LL (3LL) treated with the indicated doses of IL-6, IL-12, and IFN-γ for one day. The data are representative of more than three independent experiments. * p <0.01.

Article Snippet: The mouse spontaneous mammary tumor (MMT) cell line MMT060562 and the Ex-3LL (3LL) mouse Lewis lung carcinoma cell line was purchased from the JCRB Cell Bank (Osaka, JP).

Techniques:

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

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Challenge in the Pathological Diagnosis of the Follicular-Patterned Thyroid Lesions

doi: 10.31557/APJCP.2021.22.10.3365

Figure Lengend Snippet: The Antibodies with Their Clones, Antigen Retrieval Methods, and Staining Patterns

Article Snippet: Positive controls included mesothelioma tissue for HBME-1 (Riera et al., 1997), Ureteric tissue for CK19 (Kasper et al., 1987), papillary thyroid carcinoma for Galectin-3 (Konstantinov et al., 1996), and Small cell lung carcinoma for CD56 antibodies (Tsang et al., 1996).

Techniques: Clone Assay, Staining, Incubation, Positive Control

Journal: Translational Cancer Research

Article Title: Chronic nicotine exposure affects programmed death-ligand 1 expression and sensitivity to epidermal growth factor receptor-tyrosine kinase inhibitor in lung cancer

doi: 10.21037/tcr.2019.05.02

Figure Lengend Snippet: Morphology of cells after treatment with gefitinib in PC9 and PC9/N cells. Cells were seeded in six-well culture plates, and after 24 hrs, the cells were treated with gefitinib (0, 0.1, or 1 µM) for 48 hrs. Cells were photographed under a phase contrast inverted microscope. Cells were stained with Wright and Giemsa (original magnification 200×). These results are representative of technical triplicate.

Article Snippet: Human NSCLC cell line PC9 (EGFR exon 19 deletion) was purchased from Korean Cell Line Bank (Seoul, Korea).

Techniques: Inverted Microscopy, Staining

Journal: Translational Cancer Research

Article Title: Chronic nicotine exposure affects programmed death-ligand 1 expression and sensitivity to epidermal growth factor receptor-tyrosine kinase inhibitor in lung cancer

doi: 10.21037/tcr.2019.05.02

Figure Lengend Snippet: Cell viability assay for PC9 and PC9/N cells cultured in the presence of gefitinib. Cell growth inhibition in response to gefitinib was evaluated by using the MTT assay. Cells were treated with the indicated concentrations of gefitinib and cell viability was determined 48 hrs later. There are mean of independent triplicate experiments. The data are presented as means ± SEM from technical triplicate. PC9/N: nicotine (1 µM) expose for 3 months in PC9 cells. *, P<0.05; **, P<0.01 and ***, P<0.001 compared with PC9 + gefitinib 0 µM. ## , P<0.01 and ### , P<0.001 compared with PC9/N + gefitinib 0 µM. $ , P<0.05 between two cells with gefitinib 0.01 µM. Data are presented as mean ± standard error of the mean (SEM). SEM, standard error.

Article Snippet: Human NSCLC cell line PC9 (EGFR exon 19 deletion) was purchased from Korean Cell Line Bank (Seoul, Korea).

Techniques: Viability Assay, Cell Culture, Inhibition, MTT Assay

Journal: Translational Cancer Research

Article Title: Chronic nicotine exposure affects programmed death-ligand 1 expression and sensitivity to epidermal growth factor receptor-tyrosine kinase inhibitor in lung cancer

doi: 10.21037/tcr.2019.05.02

Figure Lengend Snippet: The mRNA expression of cells after treatment with gefitinib in PC9 and PC9/N cells. Cells were treated with gefitinib (0, 0.1, or 1 µM) for 48 hrs, and the mRNA expressions of α1-nAchR, and PD-L1 were examined by quantitative reverse transcription (qRT)-PCR (A) and RT-PCR (B). These results are representative of technical triplicate. *, P<0.05 and ***, P<0.001 compared with PC9 cells. ### , P<0.001 compared with PC9 cells + gefitinib 0 µM and $$$ , P<0.001 compared with PC9/N + gefitinib 0 µM. nAchR, nicotinic acetylcholine receptors; PD-L1, programmed death ligand 1.

Article Snippet: Human NSCLC cell line PC9 (EGFR exon 19 deletion) was purchased from Korean Cell Line Bank (Seoul, Korea).

Techniques: Expressing, Reverse Transcription, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction

Journal: Translational Cancer Research

Article Title: Chronic nicotine exposure affects programmed death-ligand 1 expression and sensitivity to epidermal growth factor receptor-tyrosine kinase inhibitor in lung cancer

doi: 10.21037/tcr.2019.05.02

Figure Lengend Snippet: The protein expression levels of cells after treatment with gefitinib in PC9 and PC9/N cells. The cells were cultured with gefitinib (0, 0.1, or 1 µM) for 48 hrs, and EGFR, mTOR, AKT, PD-L1 and α1-nAchR was detected by Western blot. β-actin was used as an internal control. Western blot was quantified by densitometry and ImageJ. These results are representative of technical triplicate. EGFR, epidermal growth factor receptor; PD-L1, programmed death ligand 1; nAchR, nicotinic acetylcholine receptors.

Article Snippet: Human NSCLC cell line PC9 (EGFR exon 19 deletion) was purchased from Korean Cell Line Bank (Seoul, Korea).

Techniques: Expressing, Cell Culture, Western Blot, Control

Journal: Translational Cancer Research

Article Title: Chronic nicotine exposure affects programmed death-ligand 1 expression and sensitivity to epidermal growth factor receptor-tyrosine kinase inhibitor in lung cancer

doi: 10.21037/tcr.2019.05.02

Figure Lengend Snippet: Immunofluorescence staining of PD-L1 and phosphorylation of EGFR in PC9 and PC9/N cells treated with 0.1 µM gefitinib. The localizations of PD-L1 and p-EGFR (green signal; Alexa488) in PC9 and PC9/N cells were shown by immunofluorescence counterstained with DAPI (blue signal) and analyzed by confocal microscopy (original magnification 400×). These results are representative of technical triplicate. DAPI, 4',6-diamidino-2-phenylindole; EGFR, epidermal growth factor receptor.

Article Snippet: Human NSCLC cell line PC9 (EGFR exon 19 deletion) was purchased from Korean Cell Line Bank (Seoul, Korea).

Techniques: Immunofluorescence, Staining, Phospho-proteomics, Confocal Microscopy

Journal: Translational Cancer Research

Article Title: Chronic nicotine exposure affects programmed death-ligand 1 expression and sensitivity to epidermal growth factor receptor-tyrosine kinase inhibitor in lung cancer

doi: 10.21037/tcr.2019.05.02

Figure Lengend Snippet: Overall proportion of PD-L1 TPS expression according to pack-year in NSCLC patients harboring activating EGFR mutation. Heavy smokers ( ≥ 30 PY) tended to have higher expression ( ≥ 50% PD-L1 TPS) than never and light smokers, however, Fisher exact test showed no statistical significance (P value =0.628). PD-L1, programmed death ligand 1; TPS, tumor proportion score; NSCLC, non-small cell lung cancer; EGFR, epidermal growth factor receptor; PY, pack-year.

Article Snippet: Human NSCLC cell line PC9 (EGFR exon 19 deletion) was purchased from Korean Cell Line Bank (Seoul, Korea).

Techniques: Expressing, Mutagenesis