cisplatin resistant lung adenocarcinoma cell line a549 ddp  (Procell Inc)

Journal: The Journal of Biological Chemistry

Article Title: Photodynamic activation of a KRAS RNA G-quadruplex–targeted photosensitizer induces ferroptosis in cisplatin-resistant non–small cell lung cancer

doi: 10.1016/j.jbc.2026.111181

Figure Lengend Snippet: Binding of MC1 to KRAS RG4 in tumor cells. A, Live A549/DDP cells stained with MC1 (2 μM), MitoTracker Red and DAPI. B, Live A549/DDP cells stained with MC1 (2 μM), with or without the pretreatment of DMS. C , Fixed A549/DDP cells stained MC1 (2 μM), with or without the pretreatment of DNase I or RNase A. D, Relative ratio of Renilla luciferase activity to Firefly luciferase activity in psiCHECK-2 vector containing mutant or wild-type 5′-UTR of KRAS, without or with the presence of MC1 . The data represent the mean ± SD (n = 5), ∗∗∗ for p < 0.001, ∗∗∗∗ for p < 0.0001 compared to the control group. RG4, RNA G-quadruplex; FL, Firefly luciferase; RL, Renilla luciferase.

Article Snippet: The authenticity of A549/DDP cell line (Procell) was validated using STR profiling, and the cell line was tested free from mycoplasma contamination.

Techniques: Binding Assay, Staining, Luciferase, Activity Assay, Plasmid Preparation, Mutagenesis, Control

Journal: The Journal of Biological Chemistry

Article Title: Photodynamic activation of a KRAS RNA G-quadruplex–targeted photosensitizer induces ferroptosis in cisplatin-resistant non–small cell lung cancer

doi: 10.1016/j.jbc.2026.111181

Figure Lengend Snippet: Antiproliferation effects of MC1 in tumor cells. A and B, transcription and expression of RAS-related genes in A549/DDP cells after 24-h treatment ( MC1 for 1 h, irradiation with 495 nm, 12.5 mW/cm 2 for 15 min, and culture for another 24 h), examined by RT-PCR and Western blot. C , ROS levels of A549/DDP cells after 24-h treatment, examined by DCFH-DA. D and E, apoptosis of A549/DDP cells after 24-h treatment, examined by Annexin V-PI staining and Western blot. F, growth inhibition of A549/DDP cells after 7-days treatment, examined by MCTS and PI staining. The data represent the mean ± SD (n = 3), ∗∗ for p < 0.01, ∗∗∗∗ for p < 0.0001 compared to the control group.

Article Snippet: The authenticity of A549/DDP cell line (Procell) was validated using STR profiling, and the cell line was tested free from mycoplasma contamination.

Techniques: Expressing, Irradiation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Staining, Inhibition, Control

Journal: The Journal of Biological Chemistry

Article Title: Photodynamic activation of a KRAS RNA G-quadruplex–targeted photosensitizer induces ferroptosis in cisplatin-resistant non–small cell lung cancer

doi: 10.1016/j.jbc.2026.111181

Figure Lengend Snippet: Ferroptosis and ICD effects of MC1 in tumor cells. A and B, GSH and NADH levels of A549/DDP cells after 24-h treatment ( MC1 for 1 h, irradiation with 495 nm, 12.5 mW/cm 2 for 15 min, and culture for another 24 h). C and D, LPO levels of A549/DDP cells after 24-h treatment, examined by flow cytometry and confocal microscopy. E, typical proteins of ferroptosis and ICD in A549/DDP cells after 24-h treatment, examined by Western blot. F, CRT expression on the surface of A549/DDP cells after 24-h treatment, examined by flow cytometry. G, extracellular ATP levels of A549/DDP cells after 24-h treatment. The data represent the mean ± SD (n = 3), ∗∗ for p < 0.01, ∗∗∗ for p < 0.001, ∗∗∗∗ for p < 0.0001 compared to the control group. ICD, immunogenic cell death; LPO, lipid peroxide; CRT, calreticulin; GSH-GPX4, glutathione peroxidase 4; HMGB1, high mobility group box 1.

Article Snippet: The authenticity of A549/DDP cell line (Procell) was validated using STR profiling, and the cell line was tested free from mycoplasma contamination.

Techniques: Irradiation, Flow Cytometry, Confocal Microscopy, Western Blot, Expressing, Control

Journal: The Journal of Biological Chemistry

Article Title: Photodynamic activation of a KRAS RNA G-quadruplex–targeted photosensitizer induces ferroptosis in cisplatin-resistant non–small cell lung cancer

doi: 10.1016/j.jbc.2026.111181

Figure Lengend Snippet: Imaging and antiproliferation effects of MC1 in vivo . A, fluorescence images of A549/DDP-bearing mice with intratumoral injection of MC1 (100 μM, 100 μl) for 0 h and 24 h, as well as the major organs and tumors of the mice harvested for 24 h. B, images of the excised tumors from A549/DDP-bearing mice treated with control, and MC1-Light (100 μM, 100 μl MC1 injection as well as irradiation with 495 nm, 12.5 mW/cm 2 for 30 min) every other day for 24 days. C , tumor growth curves displaying the tumor volumes measured every other day until 24 days after tumor implantation. D, tumor weights determined at the time of sacrifice. The data represent the mean ± SD (n = 4), ∗∗ for p < 0.01, ∗∗∗∗ for p < 0.0001 compared to the control group.

Article Snippet: The authenticity of A549/DDP cell line (Procell) was validated using STR profiling, and the cell line was tested free from mycoplasma contamination.

Techniques: Imaging, In Vivo, Fluorescence, Injection, Control, Irradiation, Tumor Implantation

Journal: Cell & Bioscience

Article Title: AFF4 promotes tumor progression and cisplatin resistance by modulating the PTEN/PI3K/AKT/mTOR axis to accelerate glycolysis in lung adenocarcinoma

doi: 10.1186/s13578-025-01455-1

Figure Lengend Snippet: AFF4 is a prognostic marker in lung adenocarcinoma (LUAD). A Kaplan–Meier analysis ( http://kmplot.com/ ) showing overall survival in LUAD patients with low and high expression of AFF4. B The differential expression of AFF4 in paired tumor tissues (tumor) and adjacent non-tumor lung tissues (normal) as shown in GSE10072 (including 58 LUAD tissues and 49 adjacent normal tissues) and GSE115002 (including 52 LUAD tissues and 52 adjacent normal tissues) datasets. C , D qPCR ( C ) and western blotting ( D ) analysis of the relative mRNA and protein levels of AFF4 in LUAD cells (A549, PC9, and H1299) and normal lung epithelial cells (Beas-2B). β-actin served as an internal control. E Quantification of AFF4 protein in Beas-2B, A549, PC9, and H1299 cells. β-actin served as an internal control. F Representative immunohistochemistry (IHC) images of AFF4 protein in LUAD tumor tissues and non-tumor tissues ( n = 8; scale bars, 100 μm). G Quantification of AFF4-positive cells. Statistical analyses were performed using the t -tests ( B , F ) and two-way ANOVA tests ( C , E ). All data are presented as the mean ± standard error of the mean; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The human embryonic kidney cell line 293T, human normal lung epithelial cell line Beas-2B, human LUAD cell lines A549, H1299, PC9, H358 and H1975, and the cisplatin-resistant cell line A549 DDP were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Marker, Expressing, Quantitative Proteomics, Western Blot, Control, Immunohistochemistry

Journal: Cell & Bioscience

Article Title: AFF4 promotes tumor progression and cisplatin resistance by modulating the PTEN/PI3K/AKT/mTOR axis to accelerate glycolysis in lung adenocarcinoma

doi: 10.1186/s13578-025-01455-1

Figure Lengend Snippet: AFF4 knockdown inhibits cell proliferation, migration, and invasion. A , B Cell counting kit-8 (CCK-8) ( A ) and clone formation assays ( B ) were used to determine cell proliferation of LUAD cells infected with control virus (NnoT sh) and two different AFF4 viruses (AFF4 sh1 and AFF4 sh2); (right) statistical analysis of violet stained cells. C Flow cytometry analysis showing the relative proportion of Ki67-positive cells in control and AFF4-deficient LUAD cells. D Representative scratch images of control and AFF4-deficient cells in 0 h compared with 36 h (PC9, H1299) or 48 h (A549); (right) calculations of mean distances. E Transwell assay detection of the cell migration and invasion abilities in NnoT sh and AFF4 sh LUAD cells; (right) record of migrated and invaded cell counts. Statistical analyses were performed using two-way ANOVA tests ( A , B , D , E ). All data are presented as the mean ± standard error of the mean; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The human embryonic kidney cell line 293T, human normal lung epithelial cell line Beas-2B, human LUAD cell lines A549, H1299, PC9, H358 and H1975, and the cisplatin-resistant cell line A549 DDP were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Knockdown, Migration, Cell Counting, CCK-8 Assay, Infection, Control, Virus, Staining, Flow Cytometry, Transwell Assay

Journal: Cell & Bioscience

Article Title: AFF4 promotes tumor progression and cisplatin resistance by modulating the PTEN/PI3K/AKT/mTOR axis to accelerate glycolysis in lung adenocarcinoma

doi: 10.1186/s13578-025-01455-1

Figure Lengend Snippet: AFF4 improves LUAD cell resistance to cisplatin. A Correlation between AFF4 expression and half-maximal inhibitory concentration (IC 50 ) of cisplatin based on the TCGA database ( n = 516) ( https://cancergenome.nih.gov ). B Relative mRNA expression of AFF4 in wild-type (WT) A549 cells and cisplatin-resistant A549 cells (A549 DDP) from GEO dataset GSE157692 ( n = 4). C Determination of IC 50 value of cisplatin in control and AFF4 overexpressed A549 and PC9 cells. D Apoptosis in A549 and PC9 cells with or without AFF4 overexpression after treatment with 10 µg/mL cisplatin for 48 h. Right panel: calculation of apoptosis cells. E Cisplatin IC 50 values in WT and A549 DDP cells. F , G AFF4 expression was detected in the WT and A549 DDP cells at mRNA ( F ) and protein levels ( G ). H Quantitative analysis of AFF4 protein. I Cisplatin IC 50 values in A549 DDP cells with or without AFF4 impairment. J Apoptosis in control and AFF4-deficient A549 DDP cells following 10 µg/mL of cisplatin treatment. Right panel: calculation of apoptosis cells. K-L Cell proliferation in AFF4-deficient A549 DDP cells as assessed using the CCK-8 ( K ) and colony formation assays ( L ); (right) violet-stained cell count. M Transwell analysis of AFF4-deficient A549 DDP cell migration and invasion. Statistical analyses were performed using t -tests ( B , C , E , F , H ) and two-way ANOVA tests ( D , I-M ). All data are presented as the mean ± standard error of the mean; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The human embryonic kidney cell line 293T, human normal lung epithelial cell line Beas-2B, human LUAD cell lines A549, H1299, PC9, H358 and H1975, and the cisplatin-resistant cell line A549 DDP were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing, Concentration Assay, Control, Over Expression, CCK-8 Assay, Staining, Cell Counting, Migration

Journal: Cell & Bioscience

Article Title: AFF4 promotes tumor progression and cisplatin resistance by modulating the PTEN/PI3K/AKT/mTOR axis to accelerate glycolysis in lung adenocarcinoma

doi: 10.1186/s13578-025-01455-1

Figure Lengend Snippet: AFF4 enhances glycolysis to promote cell proliferation, migration, and cisplatin resistance. A Gene set enrichment analysis (GSEA) of transcriptome sequencing data. B , C Glucose uptake ( B ) and lactate production ( C ) in A549, PC9, H1299 and A549 DDP cells with or without AFF4 silencing. D Representative images of reduced glucose consumption in A549, PC9, H1299 and A549 DDP cells after AFF4 silencing; 2-NBDG was added to cells at a final concentration of 100 µM for 30 min (scale bars, 50 μm). E Relative fluorescence level of 2-NBDG. F , G Glucose uptake ( F ) and lactate production ( G ) in control and AFF4-overexpressing LUAD cells (A549 and PC9) treated with vehicle or 2-deoxy-D-glucose (2-DG); cells were treated with 10 mM 2-DG for 48 h. H , I CCK-8 ( H ) and transwell assay ( I ) analyses of the proliferation, migration, and invasion of control and AFF4-overexpressing LUAD cells (A549 and PC9) with or without 2-DG treatment; (right) calculated migrated and invaded cell counts. Statistical analyses were performed using two-way ANOVA tests ( B , C , E , F–I ). All data are presented as the mean ± standard error of the mean; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The human embryonic kidney cell line 293T, human normal lung epithelial cell line Beas-2B, human LUAD cell lines A549, H1299, PC9, H358 and H1975, and the cisplatin-resistant cell line A549 DDP were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Migration, Sequencing, Concentration Assay, Fluorescence, Control, CCK-8 Assay, Transwell Assay

Journal: Cell & Bioscience

Article Title: AFF4 promotes tumor progression and cisplatin resistance by modulating the PTEN/PI3K/AKT/mTOR axis to accelerate glycolysis in lung adenocarcinoma

doi: 10.1186/s13578-025-01455-1

Figure Lengend Snippet: AFF4 promotes aerobic glycolysis by activating the PI3K/AKT/mTOR signaling pathway. A The PI3K/AKT/mTOR and mTOR1 signaling pathways were enriched in the GSEA of the AFF4 transcriptome sequencing data. B Western blotting analysis of PI3K, p-PI3K AKT, p-AKT, mTOR, p-mTOR and HIF1α abundance in control and AFF4-deficient LUAD cells (A549 and PC9). C Quantification of PI3K, p-PI3K AKT, p-AKT, mTOR, p-mTOR and HIF1α proteins. D , E Glycolysis metabolites glucose ( D ) and lactate ( E ) in control and AFF4-overexpressing A549 and PC9 cells treated with vehicle or artemisinin (ART; p-AKT inhibitor); A549 and PC9 were treated with 50µM or 100 µM artemisinin, respectively, for 48 h. F , G Effects of artemisinin on the proliferation (CCK-8 assay, F ), migration, and invasion (transwell assay, G ) of AFF4-overexpressing A549 and PC9 cells treated with vehicle or artemisinin; (right) migrated and invaded cell counts. Statistical comparisons were performed using two-way ANOVA tests ( C–G ). All data are presented as the mean ± standard error of the mean; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The human embryonic kidney cell line 293T, human normal lung epithelial cell line Beas-2B, human LUAD cell lines A549, H1299, PC9, H358 and H1975, and the cisplatin-resistant cell line A549 DDP were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Protein-Protein interactions, Sequencing, Western Blot, Control, CCK-8 Assay, Migration, Transwell Assay

Journal: Cell & Bioscience

Article Title: AFF4 promotes tumor progression and cisplatin resistance by modulating the PTEN/PI3K/AKT/mTOR axis to accelerate glycolysis in lung adenocarcinoma

doi: 10.1186/s13578-025-01455-1

Figure Lengend Snippet: AFF4 activates the PI3K/AKT/mTOR signaling pathway via transcriptional inhibition of PTEN. A Top 20 upregulated genes related to the PI3K/AKT/mTOR signaling pathway from GSEA. B PTEN expression in LUAD and normal tissues in the GSE115002 and GSE10072 datasets. C , D The mRNA ( C ) and protein levels ( D ) of PTEN in A549 and PC9 cells expressing NonT sh and AFF4 sh. E Quantification of AFF4 and PTEN proteins. F Gene tracks of CUT&Tag sequencing data in A549 and PC9 cells showing enrichment of AFF4 in the PTEN promoter site. G CUT&RUN-qPCR analysis of AFF4 binding to the PTEN promoter region, as shown by relatively higher enrichment of the anti-AFF4 than anti-IgG antibodies. H Schematic images of three predicted AFF4-binding motifs and mutant sites (Mut1, Mut2, and Mut3) at the PTEN promoter region. I Luciferase reporter plasmids with WT and three mutant PTEN promoters transfected into 293T cells expressing control or AFF4 to assess PTEN promoter activity. J , K Level of glucose uptake ( J ) and lactate production ( K ) in AFF4-deficient A549 and PC9 cells following PTEN knockdown. L CCK-8 assays were used to evaluate the effects of PTEN silencing on cell viability regulated by AFF4 overexpression. M Migration and invasion (transwell assay) of AFF4-deficient LUAD cells after PTEN impairment; (right) statistical analysis of migrated and invaded cells. Statistical analysis were performed using t -tests ( B , G ) and two-way ANOVA tests ( C , E , I–M ). All data are presented as the mean ± standard error of the mean; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The human embryonic kidney cell line 293T, human normal lung epithelial cell line Beas-2B, human LUAD cell lines A549, H1299, PC9, H358 and H1975, and the cisplatin-resistant cell line A549 DDP were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Inhibition, Expressing, Sequencing, Binding Assay, Mutagenesis, Luciferase, Transfection, Control, Activity Assay, Knockdown, CCK-8 Assay, Over Expression, Migration, Transwell Assay

Journal: Cell & Bioscience

Article Title: AFF4 promotes tumor progression and cisplatin resistance by modulating the PTEN/PI3K/AKT/mTOR axis to accelerate glycolysis in lung adenocarcinoma

doi: 10.1186/s13578-025-01455-1

Figure Lengend Snippet: YY1 binds to the AFF4 promoter to activate its expression. A Venn diagram of JASPAR, GTRD, TFDB and PROMO, identifying YY1 as a potential transcription factor of AFF4. B YY1 expression in LUAD tumor and paired non-tumor tissues in the GSE115002 and GSE10072 datasets. C , D Changes in AFF4 expression at the mRNA ( C ) and protein levels ( D ) after YY1 knockdown. E Quantification of YY1 and AFF4 proteins. F ChIP-seq tracks showing the binding of YY1 to the AFF4 promoter region. G Enrichment of YY1 in the AFF4 promoter determined by CUT&RUN-qPCR. H Three potential YY1 binding sites and mutants in the AFF4 promoter. I Luciferase reporter assay showing the activity of the AFF4 promoter in 293T cells with control or YY1 overexpression. J Western blot analysis of YY1 deficiency and AFF4 overexpression efficacy in A549 and PC9 cells. K Quantification of YY1 and AFF4 proteins. L , M The glycolytic metabolite glucose ( L ) and lactate detection ( M ) in YY1-deficient A549 and PC9 cells with or without AFF4 overexpression. N , O CCK-8 ( N ) and transwell ( O ) assay analysis of the influence of AFF4 overexpression on cell proliferation, migration, and invasion in YY1-deficient A549 and PC9 cells; (right) migrated and invaded cell counts. Statistical analyses were performed with t -tests ( B , C , E , G ) and two-way ANOVA tests ( I , K-O ). All data are presented as the mean ± standard error of the mean; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The human embryonic kidney cell line 293T, human normal lung epithelial cell line Beas-2B, human LUAD cell lines A549, H1299, PC9, H358 and H1975, and the cisplatin-resistant cell line A549 DDP were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing, Knockdown, ChIP-sequencing, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Control, Over Expression, Western Blot, CCK-8 Assay, Migration

Journal: Cell & Bioscience

Article Title: AFF4 promotes tumor progression and cisplatin resistance by modulating the PTEN/PI3K/AKT/mTOR axis to accelerate glycolysis in lung adenocarcinoma

doi: 10.1186/s13578-025-01455-1

Figure Lengend Snippet: Role of AFF4 in vivo. A Images of xenografts from mice subcutaneously injected with control or AFF4-deficient A549 cells ( n = 4). B , C Tumor volume ( B ) and tumor weights ( C ) of the control and AFF4-deficient xenograft mice. D Western blot analysis of AFF4 and PTEN abundance in xenograft mice. E Quantification of AFF4 and PTEN proteins. F IHC analysis of AFF4, PTEN, Ki67, E-cadherin, vimentin, LDHA, and PGK1 expression in AFF4-deficient xenograft mice (scale bars, 100 μm). G Statistical analysis of positive-stained cells. H Representative images of lung metastatic nodes in xenograft mice administered control or AFF4-deficient A549 cells ( n = 4). I Representative hematoxylin/eosin staining of lung metastatic nodes in xenograft mice injected with control or AFF4-deficient A549 cells (scale bars, 100 μm). J Representative IHC images of E-cadherin- and vimentin-positive cells in control and AFF4-deficient xenograft mice (scale bars, 100 μm). K Quantification of positive-stained cells. L Schematic model of the potential mechanism by which AFF4 regulates glycolysis, tumor progression, and chemoresistance. Created with BioRender ( https://www.biorender.com/ ). Statistical analyses were performed using two-way ANOVA tests ( B , C , E , G , K ). All data are presented as the mean ± standard error of the mean; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The human embryonic kidney cell line 293T, human normal lung epithelial cell line Beas-2B, human LUAD cell lines A549, H1299, PC9, H358 and H1975, and the cisplatin-resistant cell line A549 DDP were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: In Vivo, Injection, Control, Western Blot, Expressing, Staining