Journal: Cells

Article Title: Frequent Germline and Somatic Single Nucleotide Variants in the Promoter Region of the Ribosomal RNA Gene in Japanese Lung Adenocarcinoma Patients

doi: 10.3390/cells9112409

Figure Lengend Snippet: SNVs around rDNA promoter region in 20 human cancer cell lines.

Article Snippet: The human LUAC cell line A549, colon adenocarcinoma cell lines HCT116 and DLD-1, gastric adenocarcinoma cell lines Kato III and MKN45, Burkitt lymphoma cell line Raji, urinary bladder carcinoma cell line T24, hepatoblastoma cell line HepG2, and cervical carcinoma cell line HeLa were obtained from the Japanese Collection of Research Bioresources (JCRB) Cell Bank (Osaka, Japan).

Techniques:

Journal: Cells

Article Title: Frequent Germline and Somatic Single Nucleotide Variants in the Promoter Region of the Ribosomal RNA Gene in Japanese Lung Adenocarcinoma Patients

doi: 10.3390/cells9112409

Figure Lengend Snippet: Recurrence-free survival stratified by ( A ) presence or absence of germline single-nucleotide variants (SNVs) at any site at positions +1 to +100 in the rDNA promoter region (presence versus wild type (WT)) in 82 lung adenocarcinoma (LUAC) patients, and ( B ) germline SNV at positions +1 to +100 in the rDNA promoter region (presence versus WT) in Stage I LUAC excluding MIA ( n = 42), and ( C ) in Stage I/II LUAC excluding MIA ( n = 60).

Article Snippet: The human LUAC cell line A549, colon adenocarcinoma cell lines HCT116 and DLD-1, gastric adenocarcinoma cell lines Kato III and MKN45, Burkitt lymphoma cell line Raji, urinary bladder carcinoma cell line T24, hepatoblastoma cell line HepG2, and cervical carcinoma cell line HeLa were obtained from the Japanese Collection of Research Bioresources (JCRB) Cell Bank (Osaka, Japan).

Techniques:

Journal: Cells

Article Title: Frequent Germline and Somatic Single Nucleotide Variants in the Promoter Region of the Ribosomal RNA Gene in Japanese Lung Adenocarcinoma Patients

doi: 10.3390/cells9112409

Figure Lengend Snippet: Overall survival stratified by ( A ) germline single-nucleotide variants (SNVs) at any site at positions +1 to +100 in the rDNA promoter region (presence versus wild type (WT)) in 82 lung adenocarcinoma (LUAC) patients, and ( B ) germline SNV at positions +1 to +100 in the rDNA promoter region (presence versus WT) in Stage I LUAC excluding MIA ( n = 42), and ( C ) in Stage I/II LUAC excluding MIA ( n = 60).

Article Snippet: The human LUAC cell line A549, colon adenocarcinoma cell lines HCT116 and DLD-1, gastric adenocarcinoma cell lines Kato III and MKN45, Burkitt lymphoma cell line Raji, urinary bladder carcinoma cell line T24, hepatoblastoma cell line HepG2, and cervical carcinoma cell line HeLa were obtained from the Japanese Collection of Research Bioresources (JCRB) Cell Bank (Osaka, Japan).

Techniques:

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: International Journal of General Medicine

Article Title: Identification of Prognostic Factors Related to Super Enhancer-Regulated ceRNA Network in Metastatic Lung Adenocarcinoma

doi: 10.2147/IJGM.S332317

Figure Lengend Snippet: Identification of super enhancers in LUAD cells. ( A ) Enhancers were ranked according to the H3K27ac signals in HSC4 and A549 cells based on GSE143653. ( B ) Overlapping analysis of SE-associated genes in HSC4 and A549 cells. ( C ) Functional distribution of the overlapping genes.

Article Snippet: One normal human bronchial epithelial cell line (BEAS-2B) and six LUAD cell lines (H460, HCC827, A549, H1299, PC9 and Calu3) were obtained from American Type Culture Collection (ATCC, VA, USA).

Techniques: Functional Assay

Journal: International Journal of General Medicine

Article Title: Identification of Prognostic Factors Related to Super Enhancer-Regulated ceRNA Network in Metastatic Lung Adenocarcinoma

doi: 10.2147/IJGM.S332317

Figure Lengend Snippet: Effect of AC074117.1 on proliferation of LUAD cells. ( A ) qRT-PCR was used to detect the relative expression of AC074117.1 in BEAS-2B, H460, HCC827, A549, H1299, PC9 and Calu3 cells. ( B ) qRT-PCR was used to measure the knockdown efficiency of AC074117.1 in A549 and H1299 cells. ( C and D ) MTT assay was performed to evaluate changes in cell proliferation following AC074117.1 silencing in A549 and H1299 cells. **P<0.01.

Article Snippet: One normal human bronchial epithelial cell line (BEAS-2B) and six LUAD cell lines (H460, HCC827, A549, H1299, PC9 and Calu3) were obtained from American Type Culture Collection (ATCC, VA, USA).

Techniques: Quantitative RT-PCR, Expressing, Knockdown, MTT Assay

Journal: International Journal of Biological Sciences

Article Title: DNMT3a promotes LUAD cell proliferation and metastasis by activating the HDAC7 signalling pathway

doi: 10.7150/ijbs.96509

Figure Lengend Snippet: DNMT3a and HDAC7 formed a positive feedback loop promotes LUAD cell progression. a Representative western blot and statistical analysis of DNMT3a, HDAC7, ZEB1 and c-Myc expression in DNMT3a overexpression LUAD cells and control cells after treatment with SGI1027 for 48 h. β-actin was used as an internal control. b Representative western blot and statistical analysis of DNMT3a, HDAC7, ZEB1 and c-Myc expression in DNMT3a-overexpression LUAD cells and control cells after treatment with SAHA for 48 h. β-actin was used as an internal control. c Representative images and statistical analysis of the colony formation assay in the indicated groups after treatment with SGI1027 and/or SAHA for 48 h. Colonies were visualized by crystal violet staining. d Representative wound healing assay images and statistical analysis in the indicated groups after treatment with SGI1027 and/or SAHA for 48 h. The migration ability was quantified as the mean scratch area at each time point. The initial scratch area (0 h) was set as 100%. Scale bars, 100 μm (inset). e Schematic diagram of the mechanism by which DNMT3a promotes tumour proliferation and metastasis by upregulating HDAC7 and further inducing the upregulation of ZEB1 and c-Myc in LUAD. * p < 0.05 vs. the LV-Control group, ** p < 0.01 vs. the LV-Control group, ## p < 0.01 vs. the LV-DNMT3a group, $$ p < 0.01 vs. the LV-Control+SGI1027+SAHA group, && p < 0.01 vs. the LV-DNMT3a+ SGI1027+SAHA group.

Article Snippet: The human LUAD cell line (A549, SK-LU-1, H1975) was obtained from the American Type Culture Collection (ATCC) and cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco), supplemented with 10% fetal bovine serum (FBS, Gibco, NY, USA), 1% (v/v) penicillin-streptomycin solution (Hyclone).

Techniques: Western Blot, Expressing, Over Expression, Control, Colony Assay, Staining, Wound Healing Assay, Migration

Journal: Frontiers in Pharmacology

Article Title: Identification of biomarkers based on ubiquitin-correlated genes for predicting immune profile and drug sensitivity in lung adenocarcinoma

doi: 10.3389/fphar.2025.1646396

Figure Lengend Snippet: Investigating HEATR1 ’s biological function in LUAD. (A) B4GALT4 , DNAJB4 , GORAB , HEATR1 , LPGAT1 , FAT1 , GAB2 , MTMR4 and TCP11L2 gene expression levels in BEAS-2B, H2228, and A549 cells were detected by qPCR. (B,C) Verification of HEATR1 knockdown’s impact. (D,E) Verification of how HEATR1 knockdown affects H2228 and A549 cell viability. (F–I) Representative pictures of H2228 and A549 cells following HEATR1 knockdown in the wound healing experiment (magnification, ×40; scale bars = 200 μm) and transwell assay (magnification, ×100; scale bars = 200 μm). All procedures were performed in triplicate. SD ± mean, *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001 are the ways in which the data are presented.

Article Snippet: Human normal lung epithelial cells (BEAS-2B, CRL-3588) and human LUAD cells (H2228, CRL-5935 and A549, CRM-CCL-185) purchased from the American Type Culture Collection were cultivated in RPMI-1640 medium (11875093, Gibco, United States) containing 10% fetal bovine serum (FBS, 10099141, Gibco, United States) and 100 U/mL of penicillin and streptomycin (15140122, Gibco, United States).

Techniques: Gene Expression, Knockdown, Transwell 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