Journal: PLoS ONE

Article Title: Importin α3/Qip1 Is Involved in Multiplication of Mutant Influenza Virus with Alanine Mutation at Amino Acid 9 Independently of Nuclear Transport Function

doi: 10.1371/journal.pone.0055765

Figure Lengend Snippet: (A) Schematic diagram and subcellular localization of the N-terminal 110-amino acid region (NP110aa) NLS mutants. The unconventional NLS (aa 3-TKGTKRSYEQM-13), overlapping bipartite NLS (aa 90-KKTGGPIYRRVDGKWRRELILYDKEEIRRIWR-121) or bipartite NLS (aa 198-KRGINDRNFWRGENGRKTR-216) are indicated in gray in the schematic showing of full length NP and NP110aa. A549 cells were transfected with plasmids expressing WT and mutant NP110aas. The nuclear and cytoplasmic localization of WT and mutant NP110aas were determined by monitoring mRFP fluorescence. Over five hundred mRFP-expressing cells were counted and classified as nuclear or cytoplasmic localization. (B) Fluorescence microscope images showing mRFP, WT NP110aa and a mutant lacking the N-terminal 13-amino acid tail region (NP14-110aa) (upper panel). Nucleus is stained with DAPI (lower panel). (C) Fluorescence microscope images showing WT and mutant NP110aas. The localization of NP was consistent with that observed using mRFP fluorescence.

Article Snippet: Madin-Darby canine kidney (MDCK) cells , human lung carcinoma A549 cells (ATCC, cat no: CCL-185), African green monkey kidney COS-7 cells , human embryonic kidney HEK-293 cells (ATCC, cat no: CRL-1573) and HEK-293T cells were cultured in Dulbecco's modified Eagle's medium (Sigma) containing Pen Strep Glutamine (PSG, Gibco) and 10% fetal bovine serum (Sigma).

Techniques: Transfection, Expressing, Mutagenesis, Fluorescence, Microscopy, Staining

Journal: PLoS ONE

Article Title: Importin α3/Qip1 Is Involved in Multiplication of Mutant Influenza Virus with Alanine Mutation at Amino Acid 9 Independently of Nuclear Transport Function

doi: 10.1371/journal.pone.0055765

Figure Lengend Snippet: (A) A549 cells were infected with the S9A NP mutant and WT NP viruses at a multiplicity of infection of 1. At 24 h post-infection, the supernatants were harvested and the virus titer was determined using plaque assays. Data represent the relative growth of the S9A mutant compared with that of the WT. (B) Mini-genome assay using the S9A NP mutant. The effect of the S9A mutation on viral transcription was determined using a mini-genome assay with the plasmid encoding the S9A NP mutant. Luciferase activity was measured 48 h after the transfection of viral protein- (PB2, PB1, PA or NP) and vNP-luc-expressing plasmids. To analyze the effect of the S9A mutation on vRNA transcription, luciferase activity was compared with that generated by the WT. Data represent the mean ± SD of measurements from three samples in each of two independent experiments (*p<0.05, by students t -test).

Article Snippet: Madin-Darby canine kidney (MDCK) cells , human lung carcinoma A549 cells (ATCC, cat no: CCL-185), African green monkey kidney COS-7 cells , human embryonic kidney HEK-293 cells (ATCC, cat no: CRL-1573) and HEK-293T cells were cultured in Dulbecco's modified Eagle's medium (Sigma) containing Pen Strep Glutamine (PSG, Gibco) and 10% fetal bovine serum (Sigma).

Techniques: Infection, Mutagenesis, Virus, Plasmid Preparation, Luciferase, Activity Assay, Transfection, Expressing, Generated

Journal: PLoS ONE

Article Title: Importin α3/Qip1 Is Involved in Multiplication of Mutant Influenza Virus with Alanine Mutation at Amino Acid 9 Independently of Nuclear Transport Function

doi: 10.1371/journal.pone.0055765

Figure Lengend Snippet: (A) HEK-293 cells were transfected with Qip1 siRNA for 24 h. Silencing of Qip1 was confirmed by Western blotting with an antibody specific for Qip1. (B) Qip1-silenced HEK-293 cells were transfected with plasmids encoding PB2, PB1, PA, WT or S9A NP and vNP-luc, and luciferase activity was measured after 48 h. Columns and error bars represent the mean ± SD of measurements from three samples in each of three independent experiments (*p<0.05, by students t -test). (C) A549 cells were transfected with Qip1 siRNA for 48 h. Silencing of Qip1 was confirmed by Western blotting with an antibody specific against Qip1. (D) Qip1-silenced A549 cells were transfected with pCAGGS encoding mRFP-Flag-tagged WT or S9A NP110aa. The localization of NP was consistent with that observed using mRFP fluorescence. (E) The Qip1-silenced A549 cells were infected with A/WSN/33 virus containing WT or S9A NP at an MOI of 0.01. The viral titer in supernatant of non-targeting siRNA transfected (triangle) or Qip1 siRNA transfected (square) cells was determined using a plaque assay. Data represent the mean ± SD of measurements from three samples in each of three independent experiments (*p<0.05, by students t -test).

Article Snippet: Madin-Darby canine kidney (MDCK) cells , human lung carcinoma A549 cells (ATCC, cat no: CCL-185), African green monkey kidney COS-7 cells , human embryonic kidney HEK-293 cells (ATCC, cat no: CRL-1573) and HEK-293T cells were cultured in Dulbecco's modified Eagle's medium (Sigma) containing Pen Strep Glutamine (PSG, Gibco) and 10% fetal bovine serum (Sigma).

Techniques: Transfection, Western Blot, Luciferase, Activity Assay, Fluorescence, Infection, Virus, Plaque Assay

Journal: Materials

Article Title: Bio-Derived Cellulose Nanofibers for the Development Under Environmentally Assessed Conditions of Cellulose/ZnO Nanohybrids with Enhanced Biocompatibility and Antimicrobial Properties

doi: 10.3390/ma19020346

Figure Lengend Snippet: Cytotoxicity assessment of cellulose nanofiber-based formulations on human epithelial cell lines. Cell viability (%) of ( A ) A549, ( B ) Caco-2, and ( C ) HaCaT cells after 24 h exposure to different concentrations (6.25–100 µg/mL) of c-CNF, a-CNF, c-CNF/ZnO, and a-CNF/ZnO. Cells treated with PBS served as the negative control, and viability is expressed as a percentage relative to control (mean ± SD, n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison test, with differences considered significant at p ≤ 0.05, indicated as *; absence of annotation indicates no significance ( p > 0.05).

Article Snippet: A549 (representative of human lung epithelium, Cat. No 300114), Caco-2 (representative of human intestinal epithelium, Cat. No 300137), and HaCaT (representative of human skin keratinocytes, Cat. No 300493) cells were purchased from Cytion GmbH (Heidelberg, Germany).

Techniques: Negative Control, Control, Comparison

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Hsa_circ_0129047 sponges miR-665 to attenuate lung adenocarcinoma progression by upregulating protein tyrosine phosphatase receptor type B

doi: 10.4196/kjpp.2023.27.2.131

Figure Lengend Snippet: (A) RT-qPCR analysis results of circ_0129047 in both LUAD and normal tissues. (B) RT-qPCR analysis of circ_0129047 in human bronchial epithelium (BEAS-2B) and LUAD cells (A549, H1975, PC9 and Calu-3). **p < 0.01 vs . BEAS-2B. (C) Loop structures of Circ_012904. (D) Subcellular localization of circ_0129047 in PC9 and Calu-3 cells. (E) Circ_012904 was resistant to RNase R digestion. **p < 0.01 vs . RNAse R. Values are presented as mean ± SD.

Article Snippet: Human bronchial epithelium (BEAS-2B, cat#: CRL-9609), LUAD cells (A549, cat#: CRM-CCL-185 derived from the lung tissue of a white, 58-year-old male with LUAD; H1975, CRL-5908 derived from the lung tissue of a nonsmoking female with LUAD; and Calu-3, cat#: HTB-55, derived from the lung tissue of a white, 25-year-old male with LUAD) were obtained from the American Type Culture Collection (ATCC), and PC9 (cat#: 90071810, derived from the lung tissue of an undifferentiated patient with LUAD) was obtained from Sigma-Aldrich.

Techniques: Quantitative RT-PCR

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Hsa_circ_0129047 sponges miR-665 to attenuate lung adenocarcinoma progression by upregulating protein tyrosine phosphatase receptor type B

doi: 10.4196/kjpp.2023.27.2.131

Figure Lengend Snippet: (A) MiR-665 is predicted as a target of circ_0129047 through CircInteractome. (B) RIP assay was performed using Ago2 antibody in LUAD cells, and the enrichment of circ_0129047 and miR-665 was detected. **p < 0.01 vs . Anti-IgG. (C) Luciferase activity of Circ_0129047 in LUAD cells transfected with miR-665 mimics, which bind to the circ 0129047 sequence. **p < 0.01 vs . mimic-NC. (D) RT-qPCR analysis of miR-665 in LUAD tissues and normal tissues. (E) RT-qPCR analysis of miR-665 in LUAD cells (PC9 and Calu-3) and normal cells (BEAS-2B). **p < 0.01 vs . BEAS-2B. (F) Pearson analysis of miR-665 expression and circ_0129047 expression in LUAD tissues. (G) circ 0129047-overexpressing vectors (OE-circ), OE-NC, miR-665 mimic (mimic), mimic-NC, and OE+mimic were transfected into PC9 and Calu-3 cells. The expression of miR-665 was measured using RT-qPCR 48 h after transfection. **p < 0.01 vs . OE-NC; ## p < 0.01 vs . mimic-NC; && p < 0.01 vs . OE+mimic. Values are presented as mean ± SD. LUAD, lung adenocarcinoma; Ago2, Argonaute 2; WT, wild type; MUT, mutant.

Article Snippet: Human bronchial epithelium (BEAS-2B, cat#: CRL-9609), LUAD cells (A549, cat#: CRM-CCL-185 derived from the lung tissue of a white, 58-year-old male with LUAD; H1975, CRL-5908 derived from the lung tissue of a nonsmoking female with LUAD; and Calu-3, cat#: HTB-55, derived from the lung tissue of a white, 25-year-old male with LUAD) were obtained from the American Type Culture Collection (ATCC), and PC9 (cat#: 90071810, derived from the lung tissue of an undifferentiated patient with LUAD) was obtained from Sigma-Aldrich.

Techniques: Luciferase, Activity Assay, Transfection, Sequencing, Quantitative RT-PCR, Expressing, Mutagenesis

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Hsa_circ_0129047 sponges miR-665 to attenuate lung adenocarcinoma progression by upregulating protein tyrosine phosphatase receptor type B

doi: 10.4196/kjpp.2023.27.2.131

Figure Lengend Snippet: (A) Predicted miR-665 interactions with PTPRB 3′UTR using miRNA target prediction software based on TargetScan. (B) Luciferase reporter gene assay of PTPRB 3′UTR after miR-665 treatment. **p < 0.01 vs . mimic-NC. (C) PTPRB mRNA expression in LUAD tissues analyzed using RT-qPCR. (D) Pearson correlation analysis was used to analyze the relationship between miR-665 and PTPRB in LUAD tissues. (E) Pearson correlation analysis was used to analyze the relationship between circ_0129047 and PTPRB in LUAD tissues. (F) PTPRB mRNA expression in LUAD cells (PC9 and Calu-3) and normal cells (BEAS-2B) using RT-qPCR. **p < 0.01 vs . BEAS-2B. (G) Western blot analysis determining PTPRB expression in LUAD cells transfected with PTPRB-overexpressing vectors (OE-PTPRB), OE-NC, miR-665 mimic (mimic), mimic-NC, OE-PTPRB+mimic. **p < 0.01 vs . OE-NC; ## p < 0.01 vs . mimic-NC; && p < 0.01 vs . OE-PTPRB+mimic. Values are presented as mean ± SD. LUAD, lung adenocarcinoma; PTPRB, protein tyrosine phosphatase receptor type B; WT, wild type; MUT, mutant.

Article Snippet: Human bronchial epithelium (BEAS-2B, cat#: CRL-9609), LUAD cells (A549, cat#: CRM-CCL-185 derived from the lung tissue of a white, 58-year-old male with LUAD; H1975, CRL-5908 derived from the lung tissue of a nonsmoking female with LUAD; and Calu-3, cat#: HTB-55, derived from the lung tissue of a white, 25-year-old male with LUAD) were obtained from the American Type Culture Collection (ATCC), and PC9 (cat#: 90071810, derived from the lung tissue of an undifferentiated patient with LUAD) was obtained from Sigma-Aldrich.

Techniques: Software, Luciferase, Reporter Gene Assay, Expressing, Quantitative RT-PCR, Western Blot, Transfection, Mutagenesis

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Hsa_circ_0129047 sponges miR-665 to attenuate lung adenocarcinoma progression by upregulating protein tyrosine phosphatase receptor type B

doi: 10.4196/kjpp.2023.27.2.131

Figure Lengend Snippet: LUAD cells were transfected with PTPRB-overexpressing vectors (OE-PTPRB), OE-NC, miR-665 mimic (mimic), mimic-NC, and OE-PTPRB+mimic. (A) Cell proliferation was analyzed using the CCK-8 assay. (B) Western blot analysis of anti-Bax and anti-Bcl-2 expression in the transfected cells. (C) Colony formation was determined using colony formation assay (×200). *p < 0.05, **p < 0.01 vs . OE-NC; # p < 0.05, ## p < 0.01 vs . mimic-NC; & p < 0.05, && p < 0.01 vs . OE-PTPRB+mimic. Values are presented as mean ± SD. LUAD, lung adenocarcinoma; OD, optical density; PTPRB, protein tyrosine phosphatase receptor type B.

Article Snippet: Human bronchial epithelium (BEAS-2B, cat#: CRL-9609), LUAD cells (A549, cat#: CRM-CCL-185 derived from the lung tissue of a white, 58-year-old male with LUAD; H1975, CRL-5908 derived from the lung tissue of a nonsmoking female with LUAD; and Calu-3, cat#: HTB-55, derived from the lung tissue of a white, 25-year-old male with LUAD) were obtained from the American Type Culture Collection (ATCC), and PC9 (cat#: 90071810, derived from the lung tissue of an undifferentiated patient with LUAD) was obtained from Sigma-Aldrich.

Techniques: Transfection, CCK-8 Assay, Western Blot, Expressing, Colony Assay

Journal: Oncology Reports

Article Title: Mitochondrial DNA mutations are involved in the acquisition of cisplatin resistance in human lung cancer A549 cells

doi: 10.3892/or.2021.8243

Figure Lengend Snippet: ACR20 cells show resistance to CDDP-induced apoptosis. (A) Cells were treated with or without 80 µM cisplatin (CDDP) for 24 h followed by Annexin V/PI staining. Representative flow cytometry profiles of A549 and ACR20 cells untreated (NT) or treated with 80 µM CDDP for 24 h. (B) Quantification of apoptosis by Annexin V binding to A549 and ACR20 cells. Bars represent mean ± SE (n=6). (C) Western blot analysis of cleaved and total caspase-3 expression levels in A549 and ACR20 cells treated as indicated.

Article Snippet: Human lung carcinoma cell line A549 (cat. # RCB0098, RRID: CVCL_0023) were provided by RIKEN BioResource Research Center through the National Bio-Resource Project of the Ministry of Education, Culture, Sports, Science and Technology (Japan).

Techniques: Staining, Flow Cytometry, Binding Assay, Western Blot, Expressing

Journal: Oncology Reports

Article Title: Mitochondrial DNA mutations are involved in the acquisition of cisplatin resistance in human lung cancer A549 cells

doi: 10.3892/or.2021.8243

Figure Lengend Snippet: ACR20 cells show an increase in the number of mtDNA copies and decrease in Complex I activity. (A) Number of mtDNA copies was determined by qPCR of MT-COI relative to the nuclear α-tubulin in A549 and ACR20 cells. (B and C) Expression of TFAM (B) and POLRMT (C) mRNA was analyzed by RT-qPCR. Bars represent mean ± SE (n=4). (D) Complex I activity of extracted mitochondria was measured using MitoCheck ® Complex I Activity Assay kit. Bars represent mean ± SE (n=4). (E) Expression of NDFUB8 (E) mRNA was analyzed by RT-qPCR. Bars represent mean ± SE (n=4). (F and G) NDUFB8 expression was analyzed by western blotting. Bars represent mean ± SE (n=3). mtDNA, mitochondrial DNA.

Article Snippet: Human lung carcinoma cell line A549 (cat. # RCB0098, RRID: CVCL_0023) were provided by RIKEN BioResource Research Center through the National Bio-Resource Project of the Ministry of Education, Culture, Sports, Science and Technology (Japan).

Techniques: Activity Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Oncology Reports

Article Title: Mitochondrial DNA mutations are involved in the acquisition of cisplatin resistance in human lung cancer A549 cells

doi: 10.3892/or.2021.8243

Figure Lengend Snippet: Identified mtDNA mutations with varying percentage levels in ACR20 cells.

Article Snippet: Human lung carcinoma cell line A549 (cat. # RCB0098, RRID: CVCL_0023) were provided by RIKEN BioResource Research Center through the National Bio-Resource Project of the Ministry of Education, Culture, Sports, Science and Technology (Japan).

Techniques: Mutagenesis

Journal: Oncology Reports

Article Title: Mitochondrial DNA mutations are involved in the acquisition of cisplatin resistance in human lung cancer A549 cells

doi: 10.3892/or.2021.8243

Figure Lengend Snippet: Mitochondrial dysfunction leads to the decreased sensitivity of ACR20 cells to CDDP. (A) A scheme showing the methods for generation of A549 cyb and ACR20 cyb cells. (B) Cells were incubated with CM-H 2 DCFDA and observed by fluorescence microscopy (EVOS ® FL). Representative images under the fluorescence microscope are shown. The number of CM-H 2 DCFDA-stained cells was counted. Bars represent mean ± SE (n=3). Scale bar, 20 µm. (C) Complex I activity of extracted mitochondria was measured using MitoCheck ® Complex I Activity Assay kit. Bars represent mean ± SE (n=4). (D-F) Expression levels of XIAP (D), c-IAP1 (E) and c-IAP2 (F) were analyzed by western blotting. Bars represent mean ± SE (n=4). (G) Cells were incubated in culture medium containing various concentrations of cisplatin (CDDP) for 72 h. Cell viability was measured using crystal violet assay. Each value represents mean ± SE (n=4). Unpaired t-test; *P<0.05, **P<0.01. IAP, inhibitors of apoptosis protein; mtDNA, mitochondrial DNA.

Article Snippet: Human lung carcinoma cell line A549 (cat. # RCB0098, RRID: CVCL_0023) were provided by RIKEN BioResource Research Center through the National Bio-Resource Project of the Ministry of Education, Culture, Sports, Science and Technology (Japan).

Techniques: Incubation, Fluorescence, Microscopy, Staining, Activity Assay, Expressing, Western Blot, Crystal Violet Assay

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: A Regulatory Axis of circ_0008193/miR-1180-3p/TRIM62 Suppresses Proliferation, Migration, Invasion, and Warburg Effect in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.12659/MSM.922900

Figure Lengend Snippet: Expression of circ_0008193 in human lung adenocarcinoma (LUAD) tumor tissues and cells. Real-time quantitative polymerase chain reaction (RT-qPCR)-measured circ_0008193 expression level in ( A ) tissues from LUAD patients ( n =53) and ( B ) LUAD cell lines. ( C, D ) RT-qPCR-compared circ_0008193 and its parent gene FAM120A levels in A549 and H1975 cells. ( E ) Kaplan-Meier analysis tested the overall survival of these LUAD patients divided into low group ( n =27) and high group ( n =26) according to circ_0008193 expression level. ( F, G ) RT-qPCR-detected circ_0008193 level in hypoxic A549 and H1975 cells. *** P <0.001 from three independent experiments.

Article Snippet: Human LUAD cell lines A549 (cat: C0016002) and H1975 (cat: C0016013) and normal human lung epithelial cell line BEAS-2B (cat: T0007001) were purchased from AddexBio (San Diego, CA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: A Regulatory Axis of circ_0008193/miR-1180-3p/TRIM62 Suppresses Proliferation, Migration, Invasion, and Warburg Effect in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.12659/MSM.922900

Figure Lengend Snippet: Effect of circ_0008193 overexpression in human lung adenocarcinoma (LUAD) cells under hypoxia in vitro . ( A ) Real-time quantitative polymerase chain reaction (RT-qPCR)-confirmed circ_0008193 level in hypoxic A549 and H1975 cells transfected with vectors carrying circ_0008193 or not (vector or circ_0008193). ( B ) 3-(4, 5-Dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide (MTT) assay evaluated cell viability; ( C, D ) special kits assessed glucose and lactate concentrations; ( E, F ) western blotting measured hexokinase II (HK2) and lactate dehydrogenase A (LDHA) expression; ( G–J ) transwell determined numbers of migrating and invading cells; ( K, L ) western blotting also measured matrix metalloproteinase 2 (MMP2) and MMP9 levels in transfected A549 and H1975 cells after hypoxia treatment for 48 h. *** P <0.001 from three independent experiments.

Article Snippet: Human LUAD cell lines A549 (cat: C0016002) and H1975 (cat: C0016013) and normal human lung epithelial cell line BEAS-2B (cat: T0007001) were purchased from AddexBio (San Diego, CA, USA).

Techniques: Over Expression, In Vitro, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Plasmid Preparation, MTT Assay, Western Blot, Expressing

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: A Regulatory Axis of circ_0008193/miR-1180-3p/TRIM62 Suppresses Proliferation, Migration, Invasion, and Warburg Effect in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.12659/MSM.922900

Figure Lengend Snippet: Relationship between circ_0008193 and micro (mi)RNA (miR)-1180-3p in human lung adenocarcinoma (LUAD) tissues and cells. ( A ) Wild type of circ_0008193 (circ_0008193-WT) was predicted to contain potential binding sites of miR-1180-3p. ( B, C ) Dual-luciferase reporter assay identified luciferase activity of vectors carrying circ_0008193-WT or mutant type circ_0008193-MUT. ( D, E ) Ribonucleic acid (RNA) immunoprecipitation validated the enriched levels of circ_0008193 and miR-1180-3p. ( F, G ) Real-time quantitative polymerase chain reaction (RT-qPCR) examined miR-1180-3p level in A549 and H1975 cells in the presence of vector or circ_0008193. RT-qPCR measured miR-1180-3p expression status in ( H ) tissues from patients ( n =53) with LUAD and ( I ) LUAD cell lines. ( J, K ) RT-qPCR detected miR-1180-3p level in hypoxic A549 and H1975 cells. *** P <0.001 from three independent experiments. Grouping p Values B miR-NC-miR-1180-3p circ_0008193WT 0.0002 circ_0008193MUT 0.7676 C miR-NC-miR-1180-3p circ_0008193 WT <0.0001 circ_0008193 MUT 0.7761 D circ_0008193 Input vs. Anti-Ago2 <0.0001 Input vs. Anti-IgG <0.0001 Anti-Ago2 vs. Anti-IgG <0.0001 miR-1180-3p Input vs. Anti-Ago2 <0.0001 Input vs. Anti-IgG <0.0001 Anti-Ago2 vs. Anti-IgG <0.0001 E circ_0008193 Input vs. Anti-Ago2 <0.0001 Input vs. Anti-IgG <0.0001 Anti-Ago2 vs. Anti-IgG <0.0001 miR-1180-3p Input vs. Anti-Ago2 <0.0001 Input vs. Anti-IgG <0.0001 Anti-Ago2 vs. Anti-IgG <0.0001 F circ_0008193 vs. vector 0.0002 G circ_0008193 vs. vector 0.0005 H Tumor vs. mormal <0.0001 I BEAS-2B vs. A549 <0.0001 BEAS-2B vs. H1975 <0.0001 J 48 h vs. 0 h <0.0001 K 48 h vs. 0 h <0.0001

Article Snippet: Human LUAD cell lines A549 (cat: C0016002) and H1975 (cat: C0016013) and normal human lung epithelial cell line BEAS-2B (cat: T0007001) were purchased from AddexBio (San Diego, CA, USA).

Techniques: Binding Assay, Luciferase, Reporter Assay, Activity Assay, Mutagenesis, RNA Immunoprecipitation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Plasmid Preparation, Expressing

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: A Regulatory Axis of circ_0008193/miR-1180-3p/TRIM62 Suppresses Proliferation, Migration, Invasion, and Warburg Effect in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.12659/MSM.922900

Figure Lengend Snippet: Impact of miR-1180-3p expression on the role of circ_0008193 in human lung adenocarcinoma (LUAD) cells under hypoxia in vitro . ( A ) Real-time quantitative polymerase chain reaction (RT-qPCR) confirmed miR-1180-3p level in hypoxic A549 and H1975 cells cotransfected with circ_0008193 and miR-1180-3p mimic (miR-1180-3p) or its control miR-NC. ( B ) 3-(4, 5-Dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide (MTT) assay evaluated cell viability; ( C, D ) special kits assessed glucose and lactate concentrations; ( E, F ) western blotting measured hexokinase II (HK2) and lactate dehydrogenase A (LDHA) expression; ( G, H ) transwell determined numbers of migrating and invading cells; ( I, J ) western blotting also measured metalloproteinase (MMP)2 and MMP9 levels in transfected A549 and H1975 cells after hypoxia treatment for 48 h. *** P <0.001 from three independent experiments.

Article Snippet: Human LUAD cell lines A549 (cat: C0016002) and H1975 (cat: C0016013) and normal human lung epithelial cell line BEAS-2B (cat: T0007001) were purchased from AddexBio (San Diego, CA, USA).

Techniques: Expressing, In Vitro, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Control, MTT Assay, Western Blot, Transfection

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: A Regulatory Axis of circ_0008193/miR-1180-3p/TRIM62 Suppresses Proliferation, Migration, Invasion, and Warburg Effect in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.12659/MSM.922900

Figure Lengend Snippet: Relationship between miR-1180-3p and tripartite motif containing 62 (TRIM62) in human lung adenocarcinoma (LUAD) tissues and cells. ( A ) Wild type of TRIM62 3′-untranslated region (TRIM62 3′-UTR-WT) was predicted to contain potential binding sites of miR-1180-3p. ( B, C ) Dual-luciferase reporter assay identified luciferase activity of vectors carrying TRIM62 3′-UTR-WT or mutant type (TRIM62 3′-UTR-MUT). ( D, E ) Western blotting measured TRIM62 expression in A549 and H1975 cells in the presence of miR-1180-3p or miR-NC. ( F ) Western blotting identified TRIM62 expression in tissues from LUAD patients ( n =53), and three representative images (N1-3 and T1-3) are shown. ( G ) Western blotting identified TRIM62 expression in LUAD cell lines. ( H, I ) Western blotting detected TRIM62 level in hypoxic A549 and H1975 cells. ( J, K ) Western blotting detected TRIM62 level in A549 and H1975 cells in the presence of vector or circ_0008193 and copresence of circ_0008193 and miR-NC or miR-1180-3p. *** P <0.001 from three independent experiments.

Article Snippet: Human LUAD cell lines A549 (cat: C0016002) and H1975 (cat: C0016013) and normal human lung epithelial cell line BEAS-2B (cat: T0007001) were purchased from AddexBio (San Diego, CA, USA).

Techniques: Binding Assay, Luciferase, Reporter Assay, Activity Assay, Mutagenesis, Western Blot, Expressing, Plasmid Preparation

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: A Regulatory Axis of circ_0008193/miR-1180-3p/TRIM62 Suppresses Proliferation, Migration, Invasion, and Warburg Effect in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.12659/MSM.922900

Figure Lengend Snippet: Impact of tripartite motif containing 62 (TRIM62) expression on the role of circ_0008193 in human lung adenocarcinoma (LUAD) cells under hypoxia in vitro . ( A ) Western blotting confirmed TRIM62 level in hypoxic A549 and H1975 cells cotransfected with circ_0008193 and small interfering ribonucleic acid (siRNA) against TRIM62 (si-TRIM62) or its control si-NC. ( B ) 3-(4, 5-Dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide (MTT) assay evaluated cell viability; ( C, D ) special kits assessed glucose and lactate concentrations; ( E, F ) western blotting measured hexokinase II (HK2) and lactate dehydrogenase A (LDHA) expression; ( G, H ) transwell determined numbers of migrating and invading cells; ( I, J ) western blotting also measured metalloproteinase (MMP)2 and MMP9 levels in transfected A549 and H1975 cells after hypoxia treatment for 48 h. * P <0.05 and *** P <0.001 from three independent experiments.

Article Snippet: Human LUAD cell lines A549 (cat: C0016002) and H1975 (cat: C0016013) and normal human lung epithelial cell line BEAS-2B (cat: T0007001) were purchased from AddexBio (San Diego, CA, USA).

Techniques: Expressing, In Vitro, Western Blot, Control, MTT Assay, Transfection

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: A Regulatory Axis of circ_0008193/miR-1180-3p/TRIM62 Suppresses Proliferation, Migration, Invasion, and Warburg Effect in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.12659/MSM.922900

Figure Lengend Snippet: Effect of circ_0008193 overexpression on growth of A549 cells in vivo . A549 cells were subcutaneously implanted in BALB/c nude mice and then intratumorally injected with circ_0008193 ( n =4) or vector ( n =4) every 3.5 days. ( A ) Tumor volume was measured every 7 days. ( B ) Tumor weight was measured on day 35. ( C ) Images of xenograft tumors are presented. ( D–F ) Real-time quantitative polymerase chain reaction (RT-qPCR) confirmed the expression of circ_0008193, miR-1180-3p, and tripartite motif containing 62 (TRIM62) messenger ribonucleic acid (mRNA) in neoplasm tissues. ( G ) Western blotting validated TRIM62 protein expression in neoplasm tissues. *** P <0.001 from three independent experiments.

Article Snippet: Human LUAD cell lines A549 (cat: C0016002) and H1975 (cat: C0016013) and normal human lung epithelial cell line BEAS-2B (cat: T0007001) were purchased from AddexBio (San Diego, CA, USA).

Techniques: Over Expression, In Vivo, Injection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Western Blot

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: A Regulatory Axis of circ_0008193/miR-1180-3p/TRIM62 Suppresses Proliferation, Migration, Invasion, and Warburg Effect in Lung Adenocarcinoma Cells Under Hypoxia

doi: 10.12659/MSM.922900

Figure Lengend Snippet:

Article Snippet: Human LUAD cell lines A549 (cat: C0016002) and H1975 (cat: C0016013) and normal human lung epithelial cell line BEAS-2B (cat: T0007001) were purchased from AddexBio (San Diego, CA, USA).

Techniques: Plasmid Preparation

Journal: Oncology Reports

Article Title: Thermal cycling?hyperthermia sensitizes non?small cell lung cancer A549 cells to EGFR tyrosine kinase inhibitor erlotinib

doi: 10.3892/or.2025.8891

Figure Lengend Snippet: Effect of Erl or TC-HT or in combination on the cell viability and morphological changes of A549 cells at 72 h. (A) MTT viability assay of A549 non-small cell lung cancer cells treated with different concentrations of Erl or in combination with moderate temperature TC-HT treatment (41.5–43.0°C). (B) MTT viability assay of IMR-90 normal lung cells treated with various concentrations of Erl or in combination with moderate temperature TC-HT treatment (41.5–43.0°C). (C) Representative images of morphological changes of A549 cells under a light microscope after treatment with 10 µM Erl, moderate temperature TC-HT treatment (41.5–43.0°C) or in combination. Scale bar, 100 µm. Data are shown as mean ± standard deviation (n=3). Statistical significance was determined using one-way ANOVA followed by Tukey's post-hoc test. ***P<0.001. Erl, erlotinib; TC-HT, thermal cycling-hyperthermia; ctrl, control.

Article Snippet: The human NSCLC cell lines A549 (cat. no. 60074) and H460 (cat. no. 60373) and the normal lung cell line IMR-90 (cat no. 60204) were purchased from the Bioresource Collection and Research Center of the Food Industry Research and Development Institute.

Techniques: MTT Viability Assay, Light Microscopy, Standard Deviation, Control

Journal: Oncology Reports

Article Title: Thermal cycling?hyperthermia sensitizes non?small cell lung cancer A549 cells to EGFR tyrosine kinase inhibitor erlotinib

doi: 10.3892/or.2025.8891

Figure Lengend Snippet: Effect of Erl combined with TC-HT on p-EGFR protein expression in A549 cells. Western blotting of p-EGFR protein expression in A549 cells treated with 10 µM Erl, moderate temperature TC-HT, and the combination treatment. The expression levels of p-EGFR were normalized to GAPDH. Each relative expression level was compared with the control and represented as fold relative to the control. Data are shown as mean ± standard deviation (n=3). Statistical significance was determined using one-way ANOVA followed by Tukey's post-hoc test. **P<0.01, ***P<0.001. Erl, erlotinib; TC-HT, thermal cycling-hyperthermia; p, phosphorylated; t, total; ctrl, control.

Article Snippet: The human NSCLC cell lines A549 (cat. no. 60074) and H460 (cat. no. 60373) and the normal lung cell line IMR-90 (cat no. 60204) were purchased from the Bioresource Collection and Research Center of the Food Industry Research and Development Institute.

Techniques: Expressing, Western Blot, Control, Standard Deviation

Journal: Oncology Reports

Article Title: Thermal cycling?hyperthermia sensitizes non?small cell lung cancer A549 cells to EGFR tyrosine kinase inhibitor erlotinib

doi: 10.3892/or.2025.8891

Figure Lengend Snippet: Effect of Erl combined with TC-HT treatment on survival- and apoptosis-related protein expression in A549 cells. The anticancer experiments were conducted in A549 cancer cells treated with 10 µM Erl, moderate temperature TC-HT and the combination treatment. Western blotting of (A) p-JNK, (B) p-Akt and t-Akt, (C) cleaved PARP and full-length PARP (D) MTH1. The protein expression levels of p-JNK, p-Akt, t-Akt and MTH1 were normalized to GAPDH, and cleaved PARP was normalized to full-length PARP. Each relative expression level was compared with the control and represented as fold relative to the control. Data are shown as mean ± standard deviation (n=3). Statistical significance was determined using one-way ANOVA followed by Tukey's post-hoc test. *P<0.05, **P<0.01, ***P<0.001. Erl, erlotinib; TC-HT, thermal cycling-hyperthermia; p, phosphorylated; t, total; PARP, poly ADP-ribose polymerase; MTH1, MutT homolog 1; ctrl, control; ns, not significant.

Article Snippet: The human NSCLC cell lines A549 (cat. no. 60074) and H460 (cat. no. 60373) and the normal lung cell line IMR-90 (cat no. 60204) were purchased from the Bioresource Collection and Research Center of the Food Industry Research and Development Institute.

Techniques: Expressing, Western Blot, Control, Standard Deviation

Journal: Oncology Reports

Article Title: Thermal cycling?hyperthermia sensitizes non?small cell lung cancer A549 cells to EGFR tyrosine kinase inhibitor erlotinib

doi: 10.3892/or.2025.8891

Figure Lengend Snippet: Effect of Erl or TC-HT or in combination on the G2/M cell cycle arrest and protein expression levels of Cdc2 and p-p38 in A549 cells. Cell cycle and protein expression level analysis were conducted in A549 cancer cells treated with 10 µM Erl, moderate temperature TC-HT (41.5–43.0°C) and the combination treatment. (A) Flow cytometry analysis on cellular DNA content profiles of each group of A549 cells. (B) Proportion of A549 cells in G0/G1, S and G2/M phases of the indicated group. Western botting of (C) Cdc2 and (D) p-p38 protein expression. The protein expression levels of p-p38 and Cdc2 were normalized to GAPDH. Each relative expression level was compared with the control and represented as fold relative to the control. Data are shown as mean ± standard deviation (n=3). Statistical significance was determined by one-way ANOVA followed by Tukey's post-hoc test. *P<0.05, ***P<0.001. Erl, erlotinib; TC-HT, thermal cycling-hyperthermia; p, phosphorylated; t, total; ctrl, control; ns, not significant..

Article Snippet: The human NSCLC cell lines A549 (cat. no. 60074) and H460 (cat. no. 60373) and the normal lung cell line IMR-90 (cat no. 60204) were purchased from the Bioresource Collection and Research Center of the Food Industry Research and Development Institute.

Techniques: Expressing, Flow Cytometry, Western Blot, Control, Standard Deviation

Journal: Oncology Reports

Article Title: Thermal cycling?hyperthermia sensitizes non?small cell lung cancer A549 cells to EGFR tyrosine kinase inhibitor erlotinib

doi: 10.3892/or.2025.8891

Figure Lengend Snippet: Effect of Erl or TC-HT or in combination on the inhibition of A549 cell colony formation and migration. (A) Wound healing assay to determine the effect of Erl, TC-HT or combination treatment on the migration ability of A549 cells. After scratch gaps were made, A549 cells were treated with 10 µM Erl, moderate temperature TC-HT or the combination treatment. Yellow lines indicate wound edges detected by ImageJ (version 1.51j8; National Institutes of Health). Magnification, ×40. (B) Colony formation assay for A549 cells treated with 10 µM Erl, moderate temperature TC-HT or the combination treatment. (C) Wound closure rate for A549 cells was determined as the percentage of the area closed after 24 h from the initial wound area, and each group was normalized to the control group and expressed as a fraction of 100. The areas of cell-free gaps were measured and quantified using ImageJ (version 1.51j8; National Institutes of Health). (D) Colony formation rate for A549 cells at 10 days after treatment with 10 µM Erl, moderate temperature TC-HT or the combination treatment. Each group was normalized to the control group and represented as a percentage. The colony counting was performed using ImageJ (version 1.51j8; National Institutes of Health). Statistical significance was determined by one-way ANOVA followed by Tukey's post-hoc test. *P<0.05, **P<0.01, ***P<0.001. Erl, erlotinib; TC-HT, thermal cycling-hyperthermia; ctrl, control.

Article Snippet: The human NSCLC cell lines A549 (cat. no. 60074) and H460 (cat. no. 60373) and the normal lung cell line IMR-90 (cat no. 60204) were purchased from the Bioresource Collection and Research Center of the Food Industry Research and Development Institute.

Techniques: Inhibition, Migration, Wound Healing Assay, Colony Assay, Control