egfr wild type luad cell lines  (ATCC)

Journal: Molecular cancer therapeutics

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

doi: 10.1158/1535-7163.MCT-20-0024

Figure Lengend Snippet: Attenuation of YAP1 expression impairs cell proliferation and increases sensitivity to anti-cancer agents in LUAD. A, Western blotting confirmed YAP1-knockdown in H1299 and H1437 cells and YAP1-overexpression in A549 cells. B, Proliferation assay: YAP1-deficient cells (H1299 YAP1-sh and H1437 YAP1-sh) showed lower proliferative ratios than did control cells, whereas YAP1-overexpressing cells (A549 YAP1-LV) proliferated more rapidly. C, Cells were treated with CDDP, and cell viability was recorded at 72 hours after treatment. H1299 and H1437 YAP1-sh cells showed greater sensitivity to CDDP, and YAP1-LV cells showed higher resistance, than did their control counterparts. D, YAP1-deficient cells treated with CDDP (2 μM or 5 μM for 72 hours) showed greater percentages of cells undergoing apoptosis than did control cells; whereas YAP1-LV cells showed substantially less apoptosis. E, Both YAP1-sh cell lines showed significantly suppressed tumor growth in athymic mice, whereas YAP1-LV cells formed tumors more aggressively than did control cells (3 mice with tumors in both flanks per each group). Error bars: mean ± SEM. *P<0.05, **P<0.001.

Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).

Techniques: Expressing, Western Blot, Knockdown, Over Expression, Proliferation Assay, Control

Journal: Molecular cancer therapeutics

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

doi: 10.1158/1535-7163.MCT-20-0024

Figure Lengend Snippet: YAP1 promotes STAT3 phosphorylation via IL-6 upregulation. A, Immunoblotting shows YAP1, STAT3 and pSTAT3 expressions in YAP1-deficient (H1299 and H1437) and YAP1-overexpressing (A549) LUAD cells. YAP1 expression was positively associated with pSTAT3 expression. B, qRT-PCR data show IL-6 mRNA expression to be positively associated with YAP1 expression in H1299, H1437 and A549 cell lines. C, Secretion of IL-6 into media was significantly reduced in YAP1-sh cells, whereas higher levels were observed in YAP1-LV cells, as measured by ELISA in cell culture supernatant. Immunoblotting of pSTAT3 and β-actin in the respective cell lines are shown below the graph. D, Addition of recombinant human IL-6 (0.1, 1 or 10 ng/ml) to H1299 and H1437 YAP1-sh cells increased pSTAT3 levels dose-dependently. E, Blocking IL-6 activity with IL-6 neutralizing antibody (1 μg/ml) inhibited pSTAT3 in a time-dependent manner without influencing total STAT3 expression in A549 YAP1-LV cells. F, ChIP-PCR assay with YAP1 antibody was conducted using H1299, H1437 and A549 cells. DNA isolation followed by PCR showed that YAP1 protein directly binds to the IL-6 gene promoter region in three LUAD cells analyzed (white arrows). “Input” indicates DNA lysate before ChIP, which was diluted to 2% for PCR reactions. Histone H3 antibody and normal rabbit IgG served as positive and negative control, respectively. G, Schematic shows YAP1 binding to IL-6 promoter and upregulating its transcription to induce STAT3 phosphorylation. Error bars: mean ± SEM. *P<0.05, **P<0.001.

Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).

Techniques: Phospho-proteomics, Western Blot, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Recombinant, Blocking Assay, Activity Assay, DNA Extraction, Negative Control, Binding Assay

Journal: Molecular cancer therapeutics

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

doi: 10.1158/1535-7163.MCT-20-0024

Figure Lengend Snippet: Positive correlation between YAP1 and pSTAT3 expressions in human LUAD tissues. A, Representative images of YAP1 and pSTAT3 immunostaining in the TMA (scale bar: 100 μm). B, IHC results indicate a statistically significant correlation between YAP1 and pSTAT3 expressions (P<0.0001, χ2 test). C, YAP1 and pSTAT3 expressions were evaluated by immunoblotting in 13 LUAD PDXs. All YAP1-expressing PDXs (CTG0162, 0178, 0502, 0848, 1309, 1762, 2017 and 2708) also expressed pSTAT3.

Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).

Techniques: Immunostaining, Western Blot, Expressing

Journal: Molecular cancer therapeutics

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

doi: 10.1158/1535-7163.MCT-20-0024

Figure Lengend Snippet: Evaluation of verteporfin and S3I-201 as inhibitors of YAP1 and STAT3, respectively, in LUAD cells. A, Verteporfin suppressed YAP1 and STAT3 expressions in H1299 and H1437 cells in a concentration-dependent manner. STAT3 monomer (black arrows) decreased as verteporfin concentration was increased, whereas high molecular weight complexes (regions surrounded by circles) increased, indicating oligomerization of STAT3. B, S3I-201 suppressed pSTAT3 in a dose-dependent manner.

Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).

Techniques: Concentration Assay, High Molecular Weight

Journal: Molecular cancer therapeutics

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

doi: 10.1158/1535-7163.MCT-20-0024

Figure Lengend Snippet: Therapeutic efficacy of verteporfin and S3I-201 in cell lines and patient-derived preclinical xenografts mouse models of human LUAD. NSG mice were subcutaneously inoculated LUAD cell lines (H1299 and H1437 cells) and PDXs that endogenously expressed YAP1 and pSTAT3. Animals were randomly divided into different treatment groups (n=5) (A-D). Treatments were started when tumors reached 200 ± 50 mm3. A, Combined verteporfin + S3I-201 significantly inhibited tumor growth in a H1299 xenograft model. Verteporfin or S3I-201 treatment alone led to inhibited growth, but combined treatment had a greater inhibitory effect. B, In H1437 xenografts, only the combination of verteporfin + S3I-201 inhibited growth significantly more than controls, whereas either agent individually did not significantly affect tumor growth. C, D, Two EGFR wild-type PDXs (CTG0162 and CTG0178) that expressed both YAP1 and pSTAT3 (Fig. 3C) were implanted. The “All” combination (CDDP + GEM + verteporfin + S3I-201) dramatically impaired growth of both PDXs. Chemo + verteporfin also significantly retarded tumor growth until Day 28 in CTG0162 (C) and Day 34 in CTG0178 (D). However, after 4–5 weeks, tumors treated with chemo + verteporfin showed tumor regrowth. E, F, Pharmacodynamic analysis of PDX tumors treated with verteporfin, S3I-201 and chemotherapy drugs. Left panel: CTG0162 (E) and CTG0178 (F) tumors resected at 18 days after treatment (one sample per treatment). Both YAP1 and STAT3 were suppressed in the chemo + verteporfin and “All” treatment groups. Adding verteporfin to chemotherapy decreased NANOG and SOX2 expressions. Right panel: tumors resected at 43 days in CTG0162 (E) and 46 days in CTG0178 (F) were assessed (two samples per treatment). Expressions of YAP1 and STAT3 were higher in tumors treated with chemo + verteporfin than with the “All” group. Tumors from the “All” group had lower expression of NANOG in CTG0162, and both NANOG and SOX2 in CTG0178, than did the chemo + verteporfin group. Error bars: mean ± SEM. *P<0.05, **P<0.001. N.S., not significant.

Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).

Techniques: Drug discovery, Derivative Assay, Expressing

Journal: Molecular Medicine Reports

Article Title: Schisantherin A induces ferroptosis in non-small cell lung cancer through activation of the YAP/ACSL4/TfR signaling pathway

doi: 10.3892/mmr.2025.13734

Figure Lengend Snippet: Lung cancer cell viability is suppressed by Sch A in a concentration- and time-dependent manner. Sch A decreased (A) A549 and (B) HCC827 cell viability in a concentration-dependent manner, according to Cell Counting Kit-8 analysis. Sch A significantly reduced the viability of (C) A549 and (D) HCC827 cells at 24, 48 and 72 h. ***P<0.001 vs. Con. Con, control; Sch A, Schisantherin A; OD, optical density.

Article Snippet: Procell Life Science & Technology Co., Ltd. provided the EGFR-mutant HCC827 cell line (human lung adenocarcinoma origin, harboring the classic EGFR Exon19del E746-A750 deletion mutation; cat. no. CL-0094) and the EGFR wild-type A549 cell line (cat. no. CL-0016).

Techniques: Concentration Assay, Cell Counting, Control

Journal: Molecular Medicine Reports

Article Title: Schisantherin A induces ferroptosis in non-small cell lung cancer through activation of the YAP/ACSL4/TfR signaling pathway

doi: 10.3892/mmr.2025.13734

Figure Lengend Snippet: Sch A induces cell death and cell cycle arrest in A549 and HCC827 cells. Sch A considerably increased the death of (A) A549 and (B) HCC827 cells in comparison with the control group, according to flow cytometric analysis. Compared with Con, (C) A549 and (D) HCC827 cells presented a significant decrease in the number of G 2 + S-phase cells and a significant increase in the number of G 1 -phase cells. Fer-1 and DFO were able to significantly reverse the Sch A-induced decrease in (E) A549 and (F) HCC827 cell viability, according to the results of the Cell Counting Kit-8 analysis. *P<0.05, **P<0.01 and ***P<0.001 vs. Con; ## P<0.01 and ### P<0.001 vs. Sch A. DFO, deferoxamine; Con, control; Sch A, Schisantherin A; Fer-1, ferrostatin-1; Nec-1, necrostatin-1; 3-MA, 3-methyladenine.

Article Snippet: Procell Life Science & Technology Co., Ltd. provided the EGFR-mutant HCC827 cell line (human lung adenocarcinoma origin, harboring the classic EGFR Exon19del E746-A750 deletion mutation; cat. no. CL-0094) and the EGFR wild-type A549 cell line (cat. no. CL-0016).

Techniques: Comparison, Control, Cell Counting

Journal: Molecular Medicine Reports

Article Title: Schisantherin A induces ferroptosis in non-small cell lung cancer through activation of the YAP/ACSL4/TfR signaling pathway

doi: 10.3892/mmr.2025.13734

Figure Lengend Snippet: In A549 and HCC827 cells, ferroptosis inhibitors prevent Sch A-induced apoptosis. Fer-1 and DFO were shown by JC-1 staining to reverse the Sch A-induced MMP reduction in (A) A549 and (B) HCC827 cells (scale bar, 30 µm). Fer-1 and DFO inhibited the Sch A-induced increase in MDA in (C) A549 and (D) HCC827 cells. Fer-1 and DFO counteracted the Sch A-induced reduction in GSH in (E) A549 and (F) HCC827 cells. Reverse transcription-quantitative PCR analysis revealed that Fer-1 and DFO reversed the Sch A-induced increase in Chac1 and ptgs2 levels in (G) A549 and (H) HCC827 cells. *P<0.05, **P<0.01 and ***P<0.001 vs. Con; # P<0.05, ## P<0.01 and ### P<0.001 vs. Sch A. Con, control; Chac1, glutathione-specific γ-glutamylcyclotransferase 1; ptgs2, prostaglandin-endoperoxide synthase 2; Fer-1, ferrostatin-1; DFO, deferoxamine; Sch A, Schisantherin A; MDA, malondialdehyde; GSH, glutathione.

Article Snippet: Procell Life Science & Technology Co., Ltd. provided the EGFR-mutant HCC827 cell line (human lung adenocarcinoma origin, harboring the classic EGFR Exon19del E746-A750 deletion mutation; cat. no. CL-0094) and the EGFR wild-type A549 cell line (cat. no. CL-0016).

Techniques: Staining, Reverse Transcription, Real-time Polymerase Chain Reaction, Control

Journal: Molecular Medicine Reports

Article Title: Schisantherin A induces ferroptosis in non-small cell lung cancer through activation of the YAP/ACSL4/TfR signaling pathway

doi: 10.3892/mmr.2025.13734

Figure Lengend Snippet: Sch A induces ferroptosis in lung cancer cells by activating YAP signaling. (A) GeneCards intersection analysis revealed 739 overlapping genes associated with both ferroptosis and NSCLC. (B) Transcription factors among the intersecting genes were analyzed to identify key node genes. Reverse-transcription-quantitative PCR analysis revealed that Sch A increased the mRNA expression levels of YAP1 in (C) A549 and (D) HCC827 cells. Western blot analysis revealed that Sch A increased the expression levels of TfR, ACSL4 and YAP in (E) A549 and (F) HCC827 cells. *P<0.05 and **P<0.01 vs. Con. Con, control; YAP, yes-associated protein; Sch A, Schisantherin A; TfR, transferrin receptor; ACSL4, acyl-CoA synthase long-chain family member 4; NSCLC, non-small cell lung cancer.

Article Snippet: Procell Life Science & Technology Co., Ltd. provided the EGFR-mutant HCC827 cell line (human lung adenocarcinoma origin, harboring the classic EGFR Exon19del E746-A750 deletion mutation; cat. no. CL-0094) and the EGFR wild-type A549 cell line (cat. no. CL-0016).

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Control

Journal: Molecular Medicine Reports

Article Title: Schisantherin A induces ferroptosis in non-small cell lung cancer through activation of the YAP/ACSL4/TfR signaling pathway

doi: 10.3892/mmr.2025.13734

Figure Lengend Snippet: Silencing of YAP reverses Sch A-induced ferroptosis. Western blot analysis showed that the expression of YAP was decreased in (A) A549 and (B) HCC827 cells. In (C) A549 and (D) HCC827 cells, si-YAP decreased the expression of YAP and downstream signaling molecules. Fe 2+ accumulation was induced by Sch A in (E) A549 and (F) HCC827 cells, and the increase in Fe 2+ was reversed by YAP silencing, as demonstrated by FerroOrange staining (scale bar, 10 µm). After silencing YAP, the Sch A-induced increase in Fe 2+ and MDA in (G) A549 and (H) HCC827 cells was reduced. *P<0.05, **P<0.01 and ***P<0.001 vs. NC; ### P<0.001 vs. Sch A. NC, negative control; YAP, yes-associated protein; Sch A, Schisantherin A; si, small interfering RNA; MDA, malondialdehyde; ACSL4, acyl-CoA synthase long-chain family member 4; TfR, transferrin receptor.

Article Snippet: Procell Life Science & Technology Co., Ltd. provided the EGFR-mutant HCC827 cell line (human lung adenocarcinoma origin, harboring the classic EGFR Exon19del E746-A750 deletion mutation; cat. no. CL-0094) and the EGFR wild-type A549 cell line (cat. no. CL-0016).

Techniques: Western Blot, Expressing, Staining, Negative Control, Small Interfering RNA

Journal: Molecular cancer therapeutics

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

doi: 10.1158/1535-7163.MCT-20-0024

Figure Lengend Snippet: Attenuation of YAP1 expression impairs cell proliferation and increases sensitivity to anti-cancer agents in LUAD. A, Western blotting confirmed YAP1-knockdown in H1299 and H1437 cells and YAP1-overexpression in A549 cells. B, Proliferation assay: YAP1-deficient cells (H1299 YAP1-sh and H1437 YAP1-sh) showed lower proliferative ratios than did control cells, whereas YAP1-overexpressing cells (A549 YAP1-LV) proliferated more rapidly. C, Cells were treated with CDDP, and cell viability was recorded at 72 hours after treatment. H1299 and H1437 YAP1-sh cells showed greater sensitivity to CDDP, and YAP1-LV cells showed higher resistance, than did their control counterparts. D, YAP1-deficient cells treated with CDDP (2 μM or 5 μM for 72 hours) showed greater percentages of cells undergoing apoptosis than did control cells; whereas YAP1-LV cells showed substantially less apoptosis. E, Both YAP1-sh cell lines showed significantly suppressed tumor growth in athymic mice, whereas YAP1-LV cells formed tumors more aggressively than did control cells (3 mice with tumors in both flanks per each group). Error bars: mean ± SEM. *P<0.05, **P<0.001.

Article Snippet: Cell Lines and Gene Modification EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).

Techniques: Expressing, Western Blot, Knockdown, Over Expression, Proliferation Assay, Control

Journal: Molecular cancer therapeutics

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

doi: 10.1158/1535-7163.MCT-20-0024

Figure Lengend Snippet: YAP1 promotes STAT3 phosphorylation via IL-6 upregulation. A, Immunoblotting shows YAP1, STAT3 and pSTAT3 expressions in YAP1-deficient (H1299 and H1437) and YAP1-overexpressing (A549) LUAD cells. YAP1 expression was positively associated with pSTAT3 expression. B, qRT-PCR data show IL-6 mRNA expression to be positively associated with YAP1 expression in H1299, H1437 and A549 cell lines. C, Secretion of IL-6 into media was significantly reduced in YAP1-sh cells, whereas higher levels were observed in YAP1-LV cells, as measured by ELISA in cell culture supernatant. Immunoblotting of pSTAT3 and β-actin in the respective cell lines are shown below the graph. D, Addition of recombinant human IL-6 (0.1, 1 or 10 ng/ml) to H1299 and H1437 YAP1-sh cells increased pSTAT3 levels dose-dependently. E, Blocking IL-6 activity with IL-6 neutralizing antibody (1 μg/ml) inhibited pSTAT3 in a time-dependent manner without influencing total STAT3 expression in A549 YAP1-LV cells. F, ChIP-PCR assay with YAP1 antibody was conducted using H1299, H1437 and A549 cells. DNA isolation followed by PCR showed that YAP1 protein directly binds to the IL-6 gene promoter region in three LUAD cells analyzed (white arrows). “Input” indicates DNA lysate before ChIP, which was diluted to 2% for PCR reactions. Histone H3 antibody and normal rabbit IgG served as positive and negative control, respectively. G, Schematic shows YAP1 binding to IL-6 promoter and upregulating its transcription to induce STAT3 phosphorylation. Error bars: mean ± SEM. *P<0.05, **P<0.001.

Article Snippet: Cell Lines and Gene Modification EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).

Techniques: Phospho-proteomics, Western Blot, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Recombinant, Blocking Assay, Activity Assay, DNA Extraction, Negative Control, Binding Assay

Journal: Molecular cancer therapeutics

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

doi: 10.1158/1535-7163.MCT-20-0024

Figure Lengend Snippet: Positive correlation between YAP1 and pSTAT3 expressions in human LUAD tissues. A, Representative images of YAP1 and pSTAT3 immunostaining in the TMA (scale bar: 100 μm). B, IHC results indicate a statistically significant correlation between YAP1 and pSTAT3 expressions (P<0.0001, χ2 test). C, YAP1 and pSTAT3 expressions were evaluated by immunoblotting in 13 LUAD PDXs. All YAP1-expressing PDXs (CTG0162, 0178, 0502, 0848, 1309, 1762, 2017 and 2708) also expressed pSTAT3.

Article Snippet: Cell Lines and Gene Modification EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).

Techniques: Immunostaining, Western Blot, Expressing

Journal: Molecular cancer therapeutics

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

doi: 10.1158/1535-7163.MCT-20-0024

Figure Lengend Snippet: Evaluation of verteporfin and S3I-201 as inhibitors of YAP1 and STAT3, respectively, in LUAD cells. A, Verteporfin suppressed YAP1 and STAT3 expressions in H1299 and H1437 cells in a concentration-dependent manner. STAT3 monomer (black arrows) decreased as verteporfin concentration was increased, whereas high molecular weight complexes (regions surrounded by circles) increased, indicating oligomerization of STAT3. B, S3I-201 suppressed pSTAT3 in a dose-dependent manner.

Article Snippet: Cell Lines and Gene Modification EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).

Techniques: Concentration Assay, High Molecular Weight

Journal: Molecular cancer therapeutics

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

doi: 10.1158/1535-7163.MCT-20-0024

Figure Lengend Snippet: Therapeutic efficacy of verteporfin and S3I-201 in cell lines and patient-derived preclinical xenografts mouse models of human LUAD. NSG mice were subcutaneously inoculated LUAD cell lines (H1299 and H1437 cells) and PDXs that endogenously expressed YAP1 and pSTAT3. Animals were randomly divided into different treatment groups (n=5) (A-D). Treatments were started when tumors reached 200 ± 50 mm3. A, Combined verteporfin + S3I-201 significantly inhibited tumor growth in a H1299 xenograft model. Verteporfin or S3I-201 treatment alone led to inhibited growth, but combined treatment had a greater inhibitory effect. B, In H1437 xenografts, only the combination of verteporfin + S3I-201 inhibited growth significantly more than controls, whereas either agent individually did not significantly affect tumor growth. C, D, Two EGFR wild-type PDXs (CTG0162 and CTG0178) that expressed both YAP1 and pSTAT3 (Fig. 3C) were implanted. The “All” combination (CDDP + GEM + verteporfin + S3I-201) dramatically impaired growth of both PDXs. Chemo + verteporfin also significantly retarded tumor growth until Day 28 in CTG0162 (C) and Day 34 in CTG0178 (D). However, after 4–5 weeks, tumors treated with chemo + verteporfin showed tumor regrowth. E, F, Pharmacodynamic analysis of PDX tumors treated with verteporfin, S3I-201 and chemotherapy drugs. Left panel: CTG0162 (E) and CTG0178 (F) tumors resected at 18 days after treatment (one sample per treatment). Both YAP1 and STAT3 were suppressed in the chemo + verteporfin and “All” treatment groups. Adding verteporfin to chemotherapy decreased NANOG and SOX2 expressions. Right panel: tumors resected at 43 days in CTG0162 (E) and 46 days in CTG0178 (F) were assessed (two samples per treatment). Expressions of YAP1 and STAT3 were higher in tumors treated with chemo + verteporfin than with the “All” group. Tumors from the “All” group had lower expression of NANOG in CTG0162, and both NANOG and SOX2 in CTG0178, than did the chemo + verteporfin group. Error bars: mean ± SEM. *P<0.05, **P<0.001. N.S., not significant.

Article Snippet: Cell Lines and Gene Modification EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).

Techniques: Drug discovery, Derivative Assay, Expressing

Journal: Oncogenesis

Article Title: Targeting PD-L1 in non-small cell lung cancer using CAR T cells

doi: 10.1038/s41389-020-00257-z

Figure Lengend Snippet: a Flow cytometry histogram of the surface antigen expression of PD-L1 in human NSCLC cell lines and BEAS-2B. b Cytotoxic activity of PD-L1-CAR T cells after 4 and 20 h of co-culture with human NSCLC cell lines and BEAS-2B. PD-L1-CAR T cells and CD19-CAR T cells were used as effector cells at various ratios of effector (E): target (T). c Secretion of cytokines analyzed by ELISA in supernatants obtained after a 20-h co-culture of effector and target cells at a 2:1 E:T ratio. Data represented technical triplicates using T cells from one donor and were shown as mean ± SEM. **** p ≤ 0.0001.

Article Snippet: Human NSCLC EGFR- wild type cell lines A549 and H1299, EGFR -mutant cell lines HCC827 (del E746-A750) and H1975 (L858R and T790M), and normal bronchial epithelial cell line (BEAS-2B) were purchased from ATCC (Manassas, VA).

Techniques: Flow Cytometry, Expressing, Activity Assay, Co-Culture Assay, Enzyme-linked Immunosorbent Assay

Journal: Oncogenesis

Article Title: Targeting PD-L1 in non-small cell lung cancer using CAR T cells

doi: 10.1038/s41389-020-00257-z

Figure Lengend Snippet: a Experimental design of the tumor xenograft model infused with PD-L1-CAR or CD19-CAR T cells. b .NSG mice were inoculated with 1.0 × 10 6 H1975-Fluc cells and infused intravenously with 5 × 10 6 PD-L1-CAR T cells or CD19-CAR T cells twice on day 7 and 0 ( n = 5 mice per group). Bioluminescence imaging was used to assess tumor growth on day 7, 14, and 28 post tumor cell inoculation. c Bioluminescence kinetics of H1975-Fluc ( n = 5 mice per group). d The percentage of PD-L1-positive cells within tumors. All cells were extracted from tumors of each treatment group on day 28 after inoculation and the expression of PD-L1 was evaluated by flow cytometry. e Representative IHC images of CD19-CAR or PD-L1-CAR T cell-treated NSCLC tumors for PD-L1 and Ki67. Scale bars, 100 µm. f Hematoxylin and eosin staining of tumors or organs on day 28. Scale bars, 200 µm. **** p ≤ 0.0001.

Article Snippet: Human NSCLC EGFR- wild type cell lines A549 and H1299, EGFR -mutant cell lines HCC827 (del E746-A750) and H1975 (L858R and T790M), and normal bronchial epithelial cell line (BEAS-2B) were purchased from ATCC (Manassas, VA).

Techniques: Imaging, Expressing, Flow Cytometry, Staining

Journal: Oncogenesis

Article Title: Targeting PD-L1 in non-small cell lung cancer using CAR T cells

doi: 10.1038/s41389-020-00257-z

Figure Lengend Snippet: a Signal intensities of PD-L1 expression in cell lines treated with 5 Gy radiation as analyzed by flow cytometry. b Percentage of PD-L1-positive cells and cell viability in A549 cells treated with different doses of radiation for 24 or 48 h. c The effect of radiation treatment on anti-tumor efficacy of PD-L1-CAR T cells at different effector (E): target (T) ratios. Data represented technical triplicates using T cells from one donor and were shown as mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001, ns not significant.

Article Snippet: Human NSCLC EGFR- wild type cell lines A549 and H1299, EGFR -mutant cell lines HCC827 (del E746-A750) and H1975 (L858R and T790M), and normal bronchial epithelial cell line (BEAS-2B) were purchased from ATCC (Manassas, VA).

Techniques: Expressing, Flow Cytometry

Journal: Oncogenesis

Article Title: Targeting PD-L1 in non-small cell lung cancer using CAR T cells

doi: 10.1038/s41389-020-00257-z

Figure Lengend Snippet: a Experimental design of tumor cell xenograft model treated with CAR T cells and/or irradiation. b Serial bioluminescence imaging of tumor progression and regression in each group ( n = 3 mice per group). c Bioluminescence kinetics of A549-Fluc ( n = 3 mice per group) in each treatment group. d Representative IHC of PD-L1 in irradiation-treated NSCLC tumors. Scale bars, 100 µm. e Representative images of CD3 IHC in PD-L1-CAR T cell-treated and irradiation-treated NSCLC tumors. Scale bars, 100 µm. f Hematoxylin and eosin staining of tumors. Scale bars, 50 µm. * p ≤ 0.05.

Article Snippet: Human NSCLC EGFR- wild type cell lines A549 and H1299, EGFR -mutant cell lines HCC827 (del E746-A750) and H1975 (L858R and T790M), and normal bronchial epithelial cell line (BEAS-2B) were purchased from ATCC (Manassas, VA).

Techniques: Irradiation, Imaging, Staining