calu 1 cell lines  (ATCC)

Journal: Frontiers in Immunology

Article Title: Identification of immune-related prognostic biomarkers in lung squamous cell carcinoma microenvironment

doi: 10.3389/fimmu.2025.1724319

Figure Lengend Snippet: Flowchart of the study illustrating the construction and analysis of the immune-related gene (IRG) prognostic signature for LUSC.

Article Snippet: The human LUSC cell lines Calu-1 and NCI-H520, and the human monocytic cell line THP-1 were obtained from the American Type Culture Collection (ATCC).

Techniques:

Journal: Frontiers in Immunology

Article Title: Identification of immune-related prognostic biomarkers in lung squamous cell carcinoma microenvironment

doi: 10.3389/fimmu.2025.1724319

Figure Lengend Snippet: Results of immune cell infiltration between high- and low-risk groups. (A, B) Histogram of immune cell infiltration. (C) Box plot of differential expression of 22 immune cells. (D) Proportion of 22 types of immune cells in LUSC patients. (E) Comparison of stromal cells in high-risk and low-risk groups. (F–L) The correlation between characteristic genes and immune cells. Significance levels indicated are based on Benjamini-Hochberg adjusted P -values (FDR). (*p < 0.05).

Article Snippet: The human LUSC cell lines Calu-1 and NCI-H520, and the human monocytic cell line THP-1 were obtained from the American Type Culture Collection (ATCC).

Techniques: Quantitative Proteomics, Comparison

Journal: Frontiers in Immunology

Article Title: Identification of immune-related prognostic biomarkers in lung squamous cell carcinoma microenvironment

doi: 10.3389/fimmu.2025.1724319

Figure Lengend Snippet: NR1D2 is downregulated in LUSC, inhibits tumor cell migration, invasion, and modulates the immune microenvironment. (A–D) Immunohistochemistry (A, C) and Western blot (B, D) analysis show significantly reduced NR1D2 expression in LUSC tissues compared to adjacent normal lung tissues (P < 0.001). Representative images (A, B) and quantification (C, D) are shown. (E–G) Efficient knockdown of NR1D2 in Calu1 and NCI-H520 cells using two independent siRNAs (si NR1D2 #1 and si NR1D2 #2) confirmed by Western blot (E, F) and qPCR (G) (P <0.001 vs. siCtrl). (H, I) Wound healing assays demonstrate significantly impaired migratory capacity in NR1D2 -silenced Calu1 and NCI-H520 cells. Representative images (H) at 0h and 24h and quantification of wound closure (I) are shown (P < 0.001 vs. siCtrl). (J–L) Transwell assays showing significantly reduced migration and invasion of Calu1 and NCI-H520 cell lines following NR1D2 silencing. Representative images (J) and quantification of migrated/invaded cells (K, L) are presented (P < 0.001 vs. siCtrl). (M–O) Flow cytometry analysis of THP-1-derived macrophages cultured in conditioned medium (CM) from NR1D2 -silenced tumor cells. Enhanced polarization towards CD206+CD163+ M2-like macrophages was observed. Representative dot plots (M) and quantification of CD206 + CD163 + macrophage following exposure to CM from NR1D2 -depleted Calu1 cells (N) and NCI-H520 cells (O) subsets are shown (**P < 0.001 vs. CM from siCtrl cells). (P, Q) ELISA assays show increased secretion of IL-10 (P) and TGF-β1 (Q) in macrophage supernatants following exposure to CM from NR1D2 -depleted tumor cells (*P < 0.05, **P < 0.01, ***P <0.001 vs. CM from siCtrl cells). All data are presented as mean ± SD. Scale bars are provided in the respective image panels.

Article Snippet: The human LUSC cell lines Calu-1 and NCI-H520, and the human monocytic cell line THP-1 were obtained from the American Type Culture Collection (ATCC).

Techniques: Migration, Immunohistochemistry, Western Blot, Expressing, Knockdown, Flow Cytometry, Derivative Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Biological Chemistry

Article Title: Nucleolar protein DCAF13 promotes non–small cell lung cancer cell proliferation via facilitating rDNA transcription and ribosome biogenesis

doi: 10.1016/j.jbc.2025.110656

Figure Lengend Snippet: Upregulation of pre-45S rRNA in NSCLC patients. A , Pre-45S rRNA was upregulated in tumor tissues obtained from NSCLC patients in the affiliated hospital of Jiaxing university. B , the average relative expression of pre-45S rRNA in paired adjacent normal and tumor tissues was compared. ∗ p < 0.05, unpaired t test. C , H1299 cells were injected subcutaneously into both flanks of nude mice (5 × 10 6 cells per mouse, n = 6 per group). The mice were divided into two groups, and each fed with water (Control) and CX-5461 (CX-5461) every 2 days. The dosage of CX-5461 is 50 mg/kg. The tumor-bearing mice were sacrificed on day 26. Representative photographs of excised tumors are shown. D , the volume of subcutaneous tumors in the control group and CX-5461 treatment group was monitored every 2 days. ∗ p < 0.05, unpaired t test. E , tumor weight was determined at the end of the experiments. ∗ p < 0.05, unpaired t test. F , RNA was extracted from the tumors in control group and CX-5461 group. The inhibition efficiency of pre-45S rRNA in the tumors was determined by qPCR. Data are mean ± SEM. G , the average relative expression of pre-45S rRNA in control and CX-5461 group tumor tissues was compared. ∗ p < 0.05, unpaired t test. H , immunohistochemical analysis of Ki67, Brdu, p-H2AX and cleaved Caspase-3 in control and CX-5461 group tumor tissues. NSCLC, non–small cell lung cancer; qPCR, quantitative real-time PCR.

Article Snippet: Human NSCLC cell lines (A549, H1299, Calu-1, and NCI-H358), normal pulmonary bronchial epithelial cells (BEAS-2B), and human embryonic kidney cells (293T) were purchased from the American Type Culture Collection.

Techniques: Expressing, Injection, Control, Inhibition, Immunohistochemical staining, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Nucleolar protein DCAF13 promotes non–small cell lung cancer cell proliferation via facilitating rDNA transcription and ribosome biogenesis

doi: 10.1016/j.jbc.2025.110656

Figure Lengend Snippet: Silence of DCAF13 decreased the pre-45S rRNA expression, ribosomal biogenesis, and protein synthesis. A , immunofluorescent staining results showing that DCAF13 localizes at the nucleolus in H1299 and A549 cells. Cells were cultured and immunostained with anti-DCAF13 antibodies ( green ) and antibodies against the nucleolar marker B23 ( red ). The scale bars represent 20 μm. B , scatter plot to assess the correlation between DCAF13 mRNA and pre-45S rRNA in NSCLC tumors (n = 18; R 2 = 0.76). C , H1299 and A549 cells were treated with negative control siRNA, Dcaf13 siRNA1 or Dcaf13 siRNA2 for 36 h. After 36 h, cellular DCAF13 level was measured by western blots ( left panel) and qPCR ( middle panel). The pre-45S rRNA expression level was measured by qPCR ( right panel). Cells treated with Act. D (5 μM, 12 h) or CX-5461 (5 μM, 12 h) were used as a positive control. ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, unpaired t test. D , cells treated with Dcaf13 siRNA2 were transfected with DCAF13 plasmids to rescue the expression of DCAF13. The expression level of DCAF13 and pre-45S rRNA was measured by qPCR. E , the polysome profiles to monitor ribosome assembly in H1299 cells were analyzed by sucrose density gradient centrifugation. The curve graph showing the polysome profiles of H1299 cells transfected with negative control siRNA or Dcaf13 siRNA2. F , western blot results showing global protein synthesis activity in wild type H1299 and A549 cells or cells treated with negative control siRNA, Dcaf13 siRNA1, Dcaf13 siRNA2, or CX-5461 (5 μM) for 36 h. Cells were incubated in DMEM medium containing 2 μM puromycin for 3 h before being harvested for western blots using anti-puromycin antibody. G , immunofluorescent staining results showing A549 cells transiently expressing Flag-DCAF13, Flag-DCAF13 SOF1△ , and Flag-DCAF13 WD40△ were stained with anti-Flag. H , western blot results showing the indicated protein Flag-DCAF13, Flag-DCAF13 SOF1△ and Flag-DCAF13 WD40△ . I , H1299 and A549 cells transiently expressing Flag-DCAF13 were treated with 5 μM Act. D for 12 h, and then were fixed and stained with anti-Flag ( green ), anti-B23 ( red ) and DAPI ( blue ). Act. D, actinomycin D; DAPI, 4′,6-diamidino-2-phenylindole; DCAF13, DDB1- and CUL4-associated factor 13; DMEM, Dulbecco's modified Eagle's medium; NSCLC, non–small cell lung cancer; qPCR, quantitative real-time PCR.

Article Snippet: Human NSCLC cell lines (A549, H1299, Calu-1, and NCI-H358), normal pulmonary bronchial epithelial cells (BEAS-2B), and human embryonic kidney cells (293T) were purchased from the American Type Culture Collection.

Techniques: Expressing, Staining, Cell Culture, Marker, Negative Control, Western Blot, Positive Control, Transfection, Gradient Centrifugation, Activity Assay, Incubation, Modification, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Nucleolar protein DCAF13 promotes non–small cell lung cancer cell proliferation via facilitating rDNA transcription and ribosome biogenesis

doi: 10.1016/j.jbc.2025.110656

Figure Lengend Snippet: DCAF13 promotes non–small cell lung cancer progression. A , DCAF13 mRNA expression level by UALCAN. B , association between DCAF13 mRNA expression and prognosis in TCGA LUAD samples by Kaplan–Meier analysis (log-rank test). C , association between DCAF13 mRNA expression and pathologic staging. D , western blot results showing the DCAF13 expression level in NSCLC adjacent paraneoplastic tissues (NT) and tumor tissues (T). E , expression levels of DCAF13 in lung cancer cell lines (A549, NCI-H358, CALU-1, and H1299) and normal pulmonary bronchial epithelial cell (BEAS-2B) were evaluated by western blot. F , deletion of DCAF13 was confirmed at the protein level in different clones of H1299 and A549 cells by immunoblotting. G , growth curve assay for D CAF13 deletion H1299 and A549 cells. Cells (100,000) were plated in six-well culture dishes and cells were counted on days 1, 2, and 3. The data (N = 3 per cell line) are plotted as mean ± SEM. ∗∗∗ p < 0.001, unpaired t test. H , DCAF13 deletion inhibits colony formation in H1299 and A549 cells. Data are presented as mean ± SEM for N = 3 per cell line. ∗∗∗ p < 0.001. I , wound healing assay was performed for evaluating the migration ability of DCAF13 deletion H1299 and A549 cells. J , the expression of several key cell metastatic signal regulators, vimentin, MMP-9, E-cadherin, slug, snail, N-cadherin, and CCT8 were examined by qPCR. ∗∗∗ p < 0.001, unpaired t test. K , Transwell migration assay with the 24-well transwell system and quantitative analysis (original magnification: × 100). L , the numbers of migrated cells were counted in five representative high-power fields per transwell plate, ∗∗ p < 0.01. DCAF13, DDB1- and CUL4-associated factor 13; LUAD, lung adenocarcinoma; NSCLC, non–small cell lung cancer; qPCR, quantitative real-time PCR.

Article Snippet: Human NSCLC cell lines (A549, H1299, Calu-1, and NCI-H358), normal pulmonary bronchial epithelial cells (BEAS-2B), and human embryonic kidney cells (293T) were purchased from the American Type Culture Collection.

Techniques: Expressing, Western Blot, Clone Assay, Wound Healing Assay, Migration, Transwell Migration Assay, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Nucleolar protein DCAF13 promotes non–small cell lung cancer cell proliferation via facilitating rDNA transcription and ribosome biogenesis

doi: 10.1016/j.jbc.2025.110656

Figure Lengend Snippet: DCAF13 deletion inhibited the NSCLC cell proliferation in vivo . A , representative photographs of tumors formed by WT and DCAF13 deletion H1299 cells in nude mice. B , tumor weight was determined at the end of the experiments. ∗ p < 0.05, unpaired t test. C , the volume of subcutaneous tumors was monitored. ∗ p < 0.05, unpaired t test. D , RNA was extracted from the tumors in WT group and DCAF13 deletion group. The expression level of pre-45S rRNA, 28S rRNA, 18S rRNA, p53 and p27 in the tumors was determined by qPCR. Data are mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, unpaired t test. E , immunohistochemical analysis of DCAF13, p-H2AX and cleaved Caspase-3 in WT and DCAF13 deletion tumor tissues. DCAF13, DDB1- and CUL4-associated factor 13; qPCR, quantitative real-time PCR.

Article Snippet: Human NSCLC cell lines (A549, H1299, Calu-1, and NCI-H358), normal pulmonary bronchial epithelial cells (BEAS-2B), and human embryonic kidney cells (293T) were purchased from the American Type Culture Collection.

Techniques: In Vivo, Expressing, Immunohistochemical staining, Real-time Polymerase Chain Reaction