human nsclc tissue array  (Novus Biologicals)

Journal: International Journal of Biological Sciences

Article Title: Downregulation of m 6 A Reader YTHDC2 Promotes the Proliferation and Migration of Malignant Lung Cells via CYLD/NF-κB Pathway

doi: 10.7150/ijbs.58514

Figure Lengend Snippet: YTHDC2 gene and protein expression is downregulated in patients with lung cancer from The Cancer Gene Atlas, Gene Expression Omnibus and Human Protein Atlas databases, as well as in CS-exposed cells. (A) Differential analysis of YTHDC2 mRNA expression in lung cancer tissues based on the Gene Expression Profiling Interactive Analysis tool. * P<0.05 vs. normal tissues. Differential analysis of YTHDC2 mRNA expression in lung cancer tissues from (B) GSE32665 and (C) GSE19188 datasets. (D) Representative IHC images showed that YTHDC2 staining was found in the cell cytoplasm in lung cancer and normal lung tissues. High expression of YTHDC2 could be found in adjacent normal tissues, while its expression was decreased in the majority of lung cancer tissues. (E) Differential analysis of YTHDC2 staining positive ratio quantitated by IHC Profiler in lung cancer tissue arrays. YTHDC2 staining positive ratio in lung cancer tissues with different (F) maximum diameter, (G) pathological stage and (H) invasion depth. (I) Relative mRNA expression level of YTHDC2 in CS-exposed cells (S10, S20 and S30) and normal BEAS-2B cells. Western blot analysis (J) and quantitative results (K) of YTHDC2 protein expression in CS-exposed cells (S10, S20 and S30) and normal BEAS-2B cells. S10, S20 and S30 represent BEAS-2B cells exposed to CS for 10, 20 and 30 passages, respectively. **P<0.01 vs. normal BEAS-2B cells. IHC, immunohistochemistry; YTHDC2, YTH domain containing 2; CS, cigarette smoke.

Article Snippet: Human lung cancer tissue arrays (cat. no. LAC-1402 and LAC-1403) were purchased from Wuhan Servicebio Technology Co., Ltd.

Techniques: Expressing, Gene Expression, Staining, Western Blot, Immunohistochemistry

Journal: International Journal of Biological Sciences

Article Title: Downregulation of m 6 A Reader YTHDC2 Promotes the Proliferation and Migration of Malignant Lung Cells via CYLD/NF-κB Pathway

doi: 10.7150/ijbs.58514

Figure Lengend Snippet: YTHDC2 mRNA expression was regulated by gene amplification. Distribution of patients with (A) LUAD and (D) LUSC with different YTHDC2 amplification status. YTHDC2 mRNA expression in (B) LUAD and (E) LUSC tissues with different YTHDC2 amplification status. Different letters (a, b, c and d) represent statistically significant group differences. Pearson's correlation analysis revealed a significant positive correlation between YTHDC2 mRNA expression and copy numbers in (C) LUAD and (F) LUSC. The line represents linear regression of data (LUAD: y=1.065x+9.177, R 2 =0.385; LUSC: y=0.965x+9.318, R 2 =0.198). (G) The Oncomine datasets for the corresponding YTHDC2 copy numbers in lung cancer were obtained with a threshold P=0.001 and ≥2 fold-change. The data in the graphic show significant downregulation (blue column) of YTHDC2 copy numbers in lung cancer versus normal tissue. The intensity of the blue color represents the respective levels of YTHDC2 copy number. (H) Copy number variation in LUAD samples with different smoking histories. Different letters (a, b, c and d) represent statistically significant group differences. (I) Copy number variation of YTHDC2 in BEAS-2B cells and cigarette smoke-exposed cells (grey block), as well as in two lung cancer cell lines (black block). The dotted line (copy number = 2) represents the copy number of the reference gene RNase P. * P<0.05, ** P<0.01 vs. BEAS-2B cells. YTHDC2, YTH domain containing 2; LUSC, lung squamous cell carcinoma; LUAD , lung adenocarcinoma.

Article Snippet: Human lung cancer tissue arrays (cat. no. LAC-1402 and LAC-1403) were purchased from Wuhan Servicebio Technology Co., Ltd.

Techniques: Expressing, Amplification, Blocking Assay

Journal: Scientific Reports

Article Title: Novel HDAC11 inhibitors suppress lung adenocarcinoma stem cell self-renewal and overcome drug resistance by suppressing Sox2

doi: 10.1038/s41598-020-61295-6

Figure Lengend Snippet: HDAC11 expression in human lung tumor tissues and cells and its correlation with patient prognosis. ( A ) Elevated HDAC11 staining is seen in NSCLC tissue and its metastatic sites as compared to the normal human lung tissue in TMA. ( B ) Quantitation of IHC performed on the TMA shows a ~2.5–3-fold increase in HDAC11 expression in human lung adenocarcinoma tissue, squamous cell carcinoma (SCC) and metastatic lung carcinoma as compared to normal lung tissue. ( C ) Quantitation of second IHC staining for HDAC11 also showed 3-fold increase in HDAC11 expression in NSCLC patient tumor tissue as compared to normal lung tissue. ( D , E ) Kaplan-Meier survival analysis for HDAC11 (Probeset 219847_at) shows poor prognosis in lung adenocarcinoma ( D ) and squamous cell carcinoma ( E ) patients with higher mRNA expression of HDAC11 in them. ( F ) HDAC11 could be detected in multiple cell lines of different histology by a western blot analysis. Similarly, Sox2 and YAP1 could also be detected in these cells. The images of the full scan western blots are provided in Supplementary Fig. .

Article Snippet: Human NSCLC tissue array (NBP2-30222, Novus Biologicals) had 59 cores including normal lung (9 cores), NSCLCs (40 cores) and metastatic NSCLCs (10 cores).

Techniques: Expressing, Staining, Quantitation Assay, Immunohistochemistry, Western Blot

Journal: Cancers

Article Title: Epigenetic Silencing of Ubiquitin Specific Protease 4 by Snail1 Contributes to Macrophage-Dependent Inflammation and Therapeutic Resistance in Lung Cancer

doi: 10.3390/cancers12010148

Figure Lengend Snippet: Downregulation of USP4 in stemness-enriched lung cancer cells. ( A ) Top panels: Stemness of mouse D121, LLC, and human H460, HCC827, and H1299 lung cancer cell lines was enriched by sphere formation. Photos show sphere cells of each cell line. ( B ) Bottom panels: Expression of stemness-associated genes in parental D121 and LLC cells and the corresponding sphere cells analyzed by RT-qPCR. Data presented as mean ± SD of three independent experiments. ** P < 0.01.

Article Snippet: A human lung cancer tissue qPCR array (TissueScan Lung Cancer Tissue qPCR Panel III) was purchased from OriGene Technologies (Rockville, MD, USA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Cancers

Article Title: Epigenetic Silencing of Ubiquitin Specific Protease 4 by Snail1 Contributes to Macrophage-Dependent Inflammation and Therapeutic Resistance in Lung Cancer

doi: 10.3390/cancers12010148

Figure Lengend Snippet: Macrophages promote inflammation, stemness, and Snail1 expression, and downregulates USP4 in lung cancer cells. ( A ) LLC lung cancer cells were co-cultured with or without bone marrow-derived macrophages (BMDMs) in 0.4 μm transwell plates as illustrated. ( B ) Expression levels of stemness-associated genes were analyzed by RT-qPCR. ( C ) Cell surface stemness marker CD117 analyzed by flow cytometry. Left panels show a set of the histograms. ( D ) Expression levels of inflammatory cytokines analyzed by RT-qPCR. ( E , F ) Expression of Snail1 and USP4 at the mRNA level ( E ) and protein level ( F ) analyzed with RT-qPCR and immunoblotting, respectively. ( G ) Inflammatory cytokines upregulate Snail1 and downregulate USP4 in lung cancer cells. D121 and H1299 were treated with TNF-α (20 ng/mL) or IL-1β (20 ng/mL) for 48 h. Data presented as mean ± SD of three independent experiments. * P < 0.05; ** P < 0.01.

Article Snippet: A human lung cancer tissue qPCR array (TissueScan Lung Cancer Tissue qPCR Panel III) was purchased from OriGene Technologies (Rockville, MD, USA).

Techniques: Expressing, Cell Culture, Derivative Assay, Quantitative RT-PCR, Marker, Flow Cytometry, Western Blot

Journal: Cancers

Article Title: Epigenetic Silencing of Ubiquitin Specific Protease 4 by Snail1 Contributes to Macrophage-Dependent Inflammation and Therapeutic Resistance in Lung Cancer

doi: 10.3390/cancers12010148

Figure Lengend Snippet: Stable USP4 knockdown increases inflammatory status in lung cancer cells. ( A ) Efficiency of USP4 knockdown in lung cancer cell lines analyzed by immunoblotting. ( B ) NF-κB activation in control and USP4 knockdown lung cancer cells as measured by flow cytometric analysis of RelA phosphorylation. Left panels show representative histograms and right panels shows the quantitation of results. ( C – E ) Expression of cytokines in control and USP4 knockdown lung cancer cells stimulated with 10 ng/mL TNF-α with/without 1 μM BMS345541 (C,D) or 0.2 μg/mL Pam3Cys4, 5 μg/mL polyI:C, 0.2 μg/mL LPS, 2 μM R847, or CpG-1826 ( E ) for 24 h as analyzed by RT-qPCR. Data presented as mean ± SD of three independent experiments. * P < 0.05; ** P < 0.01.

Article Snippet: A human lung cancer tissue qPCR array (TissueScan Lung Cancer Tissue qPCR Panel III) was purchased from OriGene Technologies (Rockville, MD, USA).

Techniques: Knockdown, Western Blot, Activation Assay, Control, Phospho-proteomics, Quantitation Assay, Expressing, Quantitative RT-PCR

Journal: Cancers

Article Title: Epigenetic Silencing of Ubiquitin Specific Protease 4 by Snail1 Contributes to Macrophage-Dependent Inflammation and Therapeutic Resistance in Lung Cancer

doi: 10.3390/cancers12010148

Figure Lengend Snippet: Stable USP4 knockdown increases the stemness, transforming ability, and therapeutic resistance of lung cancer cells. ( A ) Expression of stemness-associated genes in control and USP4 knockdown lung cancer cells analyzed by RT-qPCR. ( B ) Control and USP4 knockdown D121 and H1299 cells were cultured in defined serum-free medium for 2 weeks to allow sphere formation. ( C ) Sphere formation compared between control D121 cells and D121 cells with stable USP4 knockdown cultured in defined serum-free medium and treated with/without 1 μM BMS345541(NF-κB inhibitor) for 2 weeks. ( D ) Control and USP4 knockdown lung cancer cells were treated with the indicated concentration of cisplatin or doxorubicin for 48 h and cell viability measured by MTS assay. ( E ) Control and USP4 knockdown lung cancer cells were treated with/without 1 μM BMS345541 for 24 h. Cell surface expression of PD-L1 as analyzed by flow cytometry. Left panels: Representative histograms. Right panels: Quantitation. Data presented as mean ± SD of three independent experiments. * P < 0.05; ** P < 0.01.

Article Snippet: A human lung cancer tissue qPCR array (TissueScan Lung Cancer Tissue qPCR Panel III) was purchased from OriGene Technologies (Rockville, MD, USA).

Techniques: Knockdown, Expressing, Control, Quantitative RT-PCR, Cell Culture, Concentration Assay, MTS Assay, Flow Cytometry, Quantitation Assay

Journal: Translational Oncology

Article Title: Vasorin/ATIA Promotes Cigarette Smoke–Induced Transformation of Human Bronchial Epithelial Cells by Suppressing Autophagy-Mediated Apoptosis

doi: 10.1016/j.tranon.2019.09.001

Figure Lengend Snippet: Increased vasorin expression in human lung cancer and HBECs exposed to tobacco carcinogens . (A) Tissue arrays were stained for vasorin with immunohistochemistry. The normal airway and alveolar epithelial cells are weakly positive while the tumor cells are strongly positive. (B) Results of tissue arrays with comparison of the staining between vasorin and normal lung epithelial cells. (C) Vasorin was examined in nontransformed (HBEC-1 and -2) and lung cancer cell lines. Relative vasorin fold increase (HBEC-2 was set to 1) was shown. (D) BEAS-2B and HBEC-2 cells were treated with CSE for 24 hours. (E) HBEC-2B (BPDE-transformed) cells were compared with HBEC-2 cells for vasorin expression. Protein was detected by western blot in C to E with β-actin as loading control.

Article Snippet: Immunohistochemistry staining using VECTASTAIN ABC Kit and DAB (3,3′-diaminobenzidine) Peroxidase Substrate Kit (Vector Laboratories, Burlingame, CA) and result assessment of human lung cancer tissue array (Imgenex; Novus Biologicals, Centennial, CO) has been described previously [ ].

Techniques: Expressing, Staining, Immunohistochemistry, Comparison, Transformation Assay, Western Blot, Control