Journal: Cells

Article Title: Dauricine Impedes the Tumorigenesis of Lung Adenocarcinoma by Regulating Nrf2 and Reactive Oxygen Species

doi: 10.3390/cells14100698

Figure Lengend Snippet: Dauricine inhibits the proliferation of NSCLC cells. Lung cancer cells ( A ) A549, ( B ) H1299, ( C ) A427, and ( D ) LLC were treated with various concentrations of dauricine (Dau) at 0, 5, 10, 15, and 20 µM for 24 h before exposed to MTT solution to assess growth progression. A colony formation assay was conducted to estimate the proliferation of lung cancer ( E , F ) A549, ( G , H ) H1299 and ( I , J ) A427 cells under the influence of dauricine at concentrations of 5 and 15 µM for 48 h. Post-treatment images were captured to perform a comparative analysis to determine the average sizes of colonies for each condition. Quantification was carried out using ImageJ, and the results are represented in columns; data are shown as mean ± SEM, with *, ** and *** indicating p < 0.05, p < 0.01 and p < 0.001, respectively. n.s., not significant.

Article Snippet: Four non-small-cell lung cancer cells were procured: A549, H1299, and Lewis Lung Carcinoma (LLC) from the American Type Culture Collection (ATCC), while A427 cells were obtained from Procell (Wuhan, China).

Techniques: Colony Assay

Journal: Cells

Article Title: Dauricine Impedes the Tumorigenesis of Lung Adenocarcinoma by Regulating Nrf2 and Reactive Oxygen Species

doi: 10.3390/cells14100698

Figure Lengend Snippet: Dauricine suppresses NSCLC cell migration. LUAD cells allowed to form a confluent monolayer were processed for uniform scratching with a 200 µL tip. ( A , B ) display H1299 cell wound recovery micrographs (Scale bar = 400 µm) of indicated concentrations and a column chart representing the quantification of wound migration. ( C , D ) A427 cell images depicting wound recovery progress along with quantification shown in column graphs for concentrations of 5 and 15 µM. Image J software was used for quantification, showing means ± SEM, with *, ** and *** indicating p < 0.05, p < 0.01 and p < 0.001 respectively.

Article Snippet: Four non-small-cell lung cancer cells were procured: A549, H1299, and Lewis Lung Carcinoma (LLC) from the American Type Culture Collection (ATCC), while A427 cells were obtained from Procell (Wuhan, China).

Techniques: Migration, Software

Journal: Cells

Article Title: Dauricine Impedes the Tumorigenesis of Lung Adenocarcinoma by Regulating Nrf2 and Reactive Oxygen Species

doi: 10.3390/cells14100698

Figure Lengend Snippet: Dauricine restrains the progression of the cell cycle and regulates cell growth. ( A ) A549, ( C ) H1299, and ( E ) A427 were exposed to doses of dauricine (5 and 15 µM) for 24 h, before EdU proliferation assay was performed as shown in the images. Post-staining images were captured using an inverted fluorescence microscope, focusing on three randomly selected fields for each condition. The quantification was analyzed for ( B ) A549, ( D ) H1299, and ( F ) A427 cells through ImageJ software and is presented in column charts. Histograms of ( G ) A549, ( I ) H1299, and ( K ) A427 illustrate the distribution of LUAD cells into distinct phases of the cell cycle after treatment with dauricine (15 µM) for 24 h. The quantification of cells in different phases is displayed on the right for ( H ) A549, ( J ) H1299, and ( L ) A427 cells. All error bars represent the standard error of the mean (n = 3), with *, ** and *** indicating p < 0.05, p < 0.01 and p < 0.001 respectively.

Article Snippet: Four non-small-cell lung cancer cells were procured: A549, H1299, and Lewis Lung Carcinoma (LLC) from the American Type Culture Collection (ATCC), while A427 cells were obtained from Procell (Wuhan, China).

Techniques: Proliferation Assay, Staining, Fluorescence, Microscopy, Software

Journal: Cells

Article Title: Dauricine Impedes the Tumorigenesis of Lung Adenocarcinoma by Regulating Nrf2 and Reactive Oxygen Species

doi: 10.3390/cells14100698

Figure Lengend Snippet: Dauricine increases the levels of ROS and reduces Nrf2 expression. ( A ) A549, ( C ) H1299, and ( E ) A427 cells were treated with dauricine at a dose of 15 µM for 24 h. Before harvesting the cells, incubation with 10 µM of DCFH-DA for 30 min was conducted to examine the ROS contents in each cell line with a flow cytometer and column charts representing the quantification of ROS in ( B ) A549, ( D ) H1299, and ( F ) A427. Cultured LUAD cells subjected to the treatment of dauricine (15 µM) with or without 10 µM of Z-VAD/fmk for 24 h were incubated with MTT solution to measure growth progression in three lung cancer cell line: ( G ) A549, ( H ) H1299, and ( I ) A427. LUAD cells were exposed to dauricine at a concentration of 15 µM for 24 h and lysates of ( J ) A549, ( L ) H1299, ( N ) A427 cells were collected to observe protein expression by Western blotting with indicated antibodies. The quantification was performed with Image Studio software and was shown in column charts for ( K ) A549, ( M ) H1299, and ( O ) A427. All error bars represent the standard error of the mean (n = 3), and * p < 0.05, ** p < 0.01.

Article Snippet: Four non-small-cell lung cancer cells were procured: A549, H1299, and Lewis Lung Carcinoma (LLC) from the American Type Culture Collection (ATCC), while A427 cells were obtained from Procell (Wuhan, China).

Techniques: Expressing, Incubation, Flow Cytometry, Cell Culture, Concentration Assay, Western Blot, Software

Journal: Cells

Article Title: Dauricine Impedes the Tumorigenesis of Lung Adenocarcinoma by Regulating Nrf2 and Reactive Oxygen Species

doi: 10.3390/cells14100698

Figure Lengend Snippet: Dauricine alters the expression of apoptosis-related proteins. ( A ) A549, ( C ) H1299, and ( E ) A427 cells were treated with 15 µM of dauricine prior to cell lysis and immunoblotting was performed with designated antibodies, and GAPDH blots display loading control. Column charts for ( B ) A549, ( D ) H1299, and ( F ) A427 represent the quantification of protein levels. All error bars represent the standard error of the mean (n = 3), and ** and *** indicating p < 0.01 and p < 0.001 respectively.

Article Snippet: Four non-small-cell lung cancer cells were procured: A549, H1299, and Lewis Lung Carcinoma (LLC) from the American Type Culture Collection (ATCC), while A427 cells were obtained from Procell (Wuhan, China).

Techniques: Expressing, Lysis, Western Blot, Control

Journal: Cancer Research

Article Title: The circRNA cEMSY Induces Immunogenic Cell Death and Boosts Immunotherapy Efficacy in Lung Adenocarcinoma

doi: 10.1158/0008-5472.CAN-24-1484

Figure Lengend Snippet: cEMSY expression level correlates with an enhanced immunotherapy response. A, Illustrative images of 65 lung adenocarcinoma (LUAD) tissues showing cEMSY (magenta), CD8 + T cells (green), CD11c + DCs (white), and DAPI (blue). Samples were categorized based on cEMSY fluorescence into cEMSY high (above the mean intensity) and cEMSY low (below the mean intensity). B, Analysis of the proportion of intratumor CD11c + DCs and CD8 + T cells within DAPI + cells across the two cEMSY stratifications. Data presented as median ± quartiles for groups cEMSY low ( n = 37) and cEMSY high ( n = 28). C, The Kaplan–Meier curve for the disease-free survival (DFS) of patients with LUAD, who were divided into two groups according to the mean expression of cEMSY. D, Heatmap showing Spearman correlations between CD11c + DCs and CD8 + T-cell infiltration and cEMSY signature gene set variation analysis score of indicated gene sets in each TCGA cohort. ACC, adrenocortical carcinoma; BLCA, bladder urothelial Carcinoma; BRCA, breast invasive carcinoma; CESC, cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL, cholangiocarcinoma; COAD, colon adenocarcinoma; DLBC, lymphoid neoplasm diffuse large B-cell lymphoma; ESCA, esophageal carcinoma; GBM, glioblastoma multiforme; HNSC, head and neck squamous cell carcinoma; KICH, kidney chromophobe; KIRC, kidney renal clear cell carcinoma; KIRP, kidney renal papillary cell carcinoma; LGG, lower grade glioma; LIHC, liver hepatocellular carcinoma; LUSC, lung squamous cell carcinoma; MESO, mesothelioma; OV, ovarian serous cystadenocarcinoma; PAAD, pancreatic adenocarcinoma; PCPG, pheochromocytoma and paraganglioma; PRAD, prostate adenocarcinoma; READ, rectum adenocarcinoma; SARC, sarcoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; TGCT, tescticular germ cell tumors; THCA, thyroid carcinoma; THYM, thymoma; UCEC, uterine corpus endometrial carcinoma; UCS, uterine carcinosarcoma; UVM, uveal melanoma. E, cEMSY signature expression level of the responders (R) and nonresponders (NR) of two representative LUAD ICB cohorts. F, Progression-free survival (PFS) of ICB cohorts based on the level of cEMSY signature. Statistics were applied using Spearman rank correlation, Student t test, or log-rank test.

Article Snippet: Human bronchial epithelial (HBE) cells and human lung adenocarcinoma cell lines (H358, A549, H23, H1975, PC9, A427, and H373), as well as murine lung adenocarcinoma cell lines (LLC1 and LA795), were obtained from the China Center for Type Culture Collection.

Techniques: Expressing, Fluorescence

Journal: EMBO Molecular Medicine

Article Title: ERK5 suppression overcomes FAK inhibitor resistance in mutant KRAS-driven non-small cell lung cancer

doi: 10.1038/s44321-024-00138-7

Figure Lengend Snippet: ( A ) Percentage of proliferation inhibition of A549, A427 and H460 cell lines treated with increasing doses of XMD8-92 or Seliciclib or in combination (top) 48 h after treatment and representative crystal violet-stained cells 72 h after drug treatment (bottom). The combination index (CI) showing the synergistic effect of combination of the 2 drugs is indicated; n = 3. ( B – D ) Relative quantification of Annexin V (AV) + Annexin V/PI (AV/PI)-positive cells by flow cytometry; n = 4 ( B ), colony forming capacity; n = 3 ( C ) and percentage of cell migration; n = 10 ( D ) of A549 cells treated with 10 µM XMD8-92 or Seliciclib or in combination, except for the colony formation assay in which cells were treated with 5 µM of each drug. ( E ) Immunoblot analysis of the indicated targets in A549 cells treated with the indicated doses of Seliciclib or XMD8-92 or in combination for 24 h. ( F ) Immunoblot analysis of the indicated targets in A549 cells treated with 10 µM XMD8-92 or 10 µM Seliciclib or in combination for 24 h. ( G ) Immunoblot analysis for the indicated targets in A549 cells transduced with shRNA control (pLKO.1 hygro + Tet-pLKO-puro) or a shRNA against CDK5 (Tet-pLKO-puro-shCDK5 + pLKO.1 hygro), or a shRNA against ERK5 (pLKO.1 hygro-shERK5 + Tet-pLKO-puro) or in combination (Tet-pLKO-puro-shCDK5 + pLKO.1 hygro-shERK5). After transduction and selection, cells were harvested for protein extraction 72 h after doxycycline (1 μg/mL) induction. ( H , I ) Relative cell number ( H ) and Annexin V (AV) + Annexin V/PI (AV/PI)-positive cell quantification by flow cytometry ( I ) in A549 cells treated as in ( G ) 72 h after doxycycline (1 μg/mL) induction; n = 3. Graphical data are mean ± SD. Statistical analyses were done using one-way ANOVA; n , number of biologically independent samples. .

Article Snippet: Human: A427 cell line , Prof. John Minna (UTSW Medical Center, Dallas, USA) , ATCC # HTB-53.

Techniques: Inhibition, Staining, Quantitative Proteomics, Flow Cytometry, Migration, Colony Assay, Western Blot, Transduction, shRNA, Control, Selection, Protein Extraction

Journal: EMBO Molecular Medicine

Article Title: ERK5 suppression overcomes FAK inhibitor resistance in mutant KRAS-driven non-small cell lung cancer

doi: 10.1038/s44321-024-00138-7

Figure Lengend Snippet: Relative quantification of cell death by flow cytometry analysis of Annexin V-Atto 633 (AV) + Annexin V/PI (AV/PI)-positive (left) and colony number (right) of A427 cells treated with XMD8-92 or Seliciclib (10 µM for apoptosis assay and 2.5 µM for colony formation each, respectively) alone or in combination; n = 3. Graphical data are mean ± SD. Statistical analyses were done using one-way ANOVA; n , number of biologically independent samples.

Article Snippet: Human: A427 cell line , Prof. John Minna (UTSW Medical Center, Dallas, USA) , ATCC # HTB-53.

Techniques: Quantitative Proteomics, Flow Cytometry, Apoptosis Assay

Journal: EMBO Molecular Medicine

Article Title: ERK5 suppression overcomes FAK inhibitor resistance in mutant KRAS-driven non-small cell lung cancer

doi: 10.1038/s44321-024-00138-7

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Human: A427 cell line , Prof. John Minna (UTSW Medical Center, Dallas, USA) , ATCC # HTB-53.

Techniques: Recombinant, Plasmid Preparation, Sequencing, shRNA, Reverse Transcription Polymerase Chain Reaction, Transfection, Western Blot, In Vitro, In Vivo, cDNA Synthesis, TUNEL Assay, Software