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: Discover oncology

Article Title: A-to-I-edited miR-1251-5p restrains tumor growth and metastasis in lung adenocarcinoma through regulating TCF7-mediated Wnt signaling pathway.

doi: 10.1007/s12672-024-01462-7

Figure Lengend Snippet: Fig. 1 MiR-1251-5p edited by ADAR1 is involved in LUAD development. A The editing level changes after transfection of wild-type ADAR enzymes (ADAR WT), and inactive ADAR enzymes (ADAR mut). B MiR-1251-5p editing in LUAD tissues and non-tumor specimens. C Dot plots showing miR-1251-5p editing in adjacent non-tumor specimens and LUAD tissues at different stage. D Kaplan-Meier plots for the dis- ease-free survival rate of LUAD patients in the groups with (+) and without (−) overediting. *P < 0.05, ** P < 0.01

Article Snippet: Human LUAD cell lines A549 and H1299 were obtained from ATCC and incubated at 37 °C with 5% CO2 in RPMI 1640 medium with 1% penicillin/streptomycin and 10% FBS.

Techniques: Transfection

Journal: Discover oncology

Article Title: A-to-I-edited miR-1251-5p restrains tumor growth and metastasis in lung adenocarcinoma through regulating TCF7-mediated Wnt signaling pathway.

doi: 10.1007/s12672-024-01462-7

Figure Lengend Snippet: Fig. 2 A-to-I-edited miR-1251-5p restrains LUAD cell viability and metastasis. A WT and ED miR-1251-5p expression was detected in A549 and H1299 cells with their mimics. B Cell proliferation detected by CCK8 in NC, WT miR-1251-5p and ED miR-1251-5p groups. C Clonal formation in NC, WT miR-1251-5p and ED miR-1251-5p groups. D Apoptosis rate in NC, WT miR-1251-5p and ED miR-1251-5p groups. E, F Cell migration and invasion detected by Wound healing and Transwell assay in NC, WT miR-1251-5p and ED miR-1251-5p groups. *P < 0.05, ** P < 0.01

Article Snippet: Human LUAD cell lines A549 and H1299 were obtained from ATCC and incubated at 37 °C with 5% CO2 in RPMI 1640 medium with 1% penicillin/streptomycin and 10% FBS.

Techniques: Expressing, Migration, Transwell Assay

Journal: Discover oncology

Article Title: A-to-I-edited miR-1251-5p restrains tumor growth and metastasis in lung adenocarcinoma through regulating TCF7-mediated Wnt signaling pathway.

doi: 10.1007/s12672-024-01462-7

Figure Lengend Snippet: Fig. 4 Edited miR-1251-5p impedes LUAD progression through disturbing Wnt signaling by suppressing TCF7. A, B The mRNA and protein expression of TCF7 in NC, TCF7 siRNA, and Si-TCF7 + ED miR-1251-5p inhibitor groups. C–G The proliferation, apoptosis, migration and inva- sion of A549 and H299 cells in NC, TCF7 siRNA, and Si-TCF7 + ED miR-1251-5p inhibitor groups. H, I CCND1 and c-MYC protein expression regulated by ED miR-1251-5p and TCF7 in LUAD cells. *P < 0.05, ** P < 0.01

Article Snippet: Human LUAD cell lines A549 and H1299 were obtained from ATCC and incubated at 37 °C with 5% CO2 in RPMI 1640 medium with 1% penicillin/streptomycin and 10% FBS.

Techniques: Expressing, Migration

Journal: Discover oncology

Article Title: A-to-I-edited miR-1251-5p restrains tumor growth and metastasis in lung adenocarcinoma through regulating TCF7-mediated Wnt signaling pathway.

doi: 10.1007/s12672-024-01462-7

Figure Lengend Snippet: Fig. 5 Edited miR-1251-5p obstructs LUAD tumor growth through disturbing Wnt signaling in vivo. A Representative tumor images. B Tumor volume in Blank, WT agomir-1251-5p and ED agomir-1251-5p groups. C Tumor weight in Blank, WT agomir-1251-5p and ED agomir- 1251-5p groups. D Representative immunohistochemistry images to analyze levels of TCF7, c-MYC, and CCND1 in tumor tissues of Blank and ED agomir-1251-5p groups. *P < 0.05, ** P < 0.01

Article Snippet: Human LUAD cell lines A549 and H1299 were obtained from ATCC and incubated at 37 °C with 5% CO2 in RPMI 1640 medium with 1% penicillin/streptomycin and 10% FBS.

Techniques: In Vivo, Immunohistochemistry

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: Frontiers in Oncology

Article Title: Pyroptosis-Related LncRNA Signatures Correlate With Lung Adenocarcinoma Prognosis

doi: 10.3389/fonc.2022.850943

Figure Lengend Snippet: (A) Expression levels of 11 pyroptosis-related lncRNAs in the normal lung epithelial cell line BEAS-2B and two LUAD cell lines (A549, NCI-H1975) by RT-qPCR. (B–K) Expression levels of 11 pyroptosis-related lncRNAs in LUAD tissues and corresponding normal tissues by RT-qPCR. Data represent the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. P values were determined by one-way ANOVA.

Article Snippet: A human LUAD cell line (A549, NCI-H1975) and a normal lung epithelial cell line (BEAS-2B) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, 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: 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: Acta biochimica et biophysica Sinica

Article Title: lncRNA CYTOR promotes lung adenocarcinoma gemcitabine resistance and epithelial-mesenchymal transition by sponging miR-125a-5p and upregulating ANLN and RRM2.

doi: 10.3724/abbs.2023287

Figure Lengend Snippet: Figure 1. CYTOR is highly expressed and acts as an unfavorable prognostic factor in LUAD (A,B) The expression of CYTOR was summarized in LUAD nonpaired subtypes (A) and paired subtypes (B) from the TCGA database. (C) The expression level of CYTOR in eight paired cancer and adjacent noncancerous tissues was examined by quantitative RT-PCR. (D) CYTOR expression was measured in LUAD and nontumor tissues using in situ hybridization technology. (E) Kaplan-Meier survival curves indicate that high expression of CYTOR is associated with poor survival in LUAD patients. (F) CYTOR expression was measured in LUAD cell lines (A549, ANIP-973, and NCI-H2122) and a human bronchial epithelial cell line (16HBE). (G–I) Comparison of CYTOR expression in LUAD patients with different T stages (T2–4 vs T1) (G), N stages (N1–3 vs T0) (H), and pathological stages (stage III & IV vs stage I & II) (I) from the TCGA LUAD subtypes. (J–L) Kaplan-Meier analysis showed that LUAD patients with high CYTOR expression had worse overall survival (J), disease-specific survival (K), and progression-free interval (L). (M) The receiver operating characteristic (ROC) curve of OS-associated CYTOR expression. Data are shown as the mean±SD, and the experiments were performed three times in triplicate. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Cell lines and patient tissues Human LUAD cell lines (A549, ANIP-973, and NCI-H2122 cells) and a bronchial epithelial cell line (16HBE cells) were obtained from ATCC (Manassas, USA).

Techniques: Expressing, Quantitative RT-PCR, In Situ Hybridization, Comparison

Journal: Acta biochimica et biophysica Sinica

Article Title: lncRNA CYTOR promotes lung adenocarcinoma gemcitabine resistance and epithelial-mesenchymal transition by sponging miR-125a-5p and upregulating ANLN and RRM2.

doi: 10.3724/abbs.2023287

Figure Lengend Snippet: Figure 2. CYTOR contributes to LUAD gemcitabine resistance and endothelial-mesenchymal transition (EMT) in vitro (A) CYTOR was overexpressed (CYTOR-overexpressing vectors) in A549 cells (left panel) or silenced (sh-CYTOR#1 and #2) in ANIP-973 cells (right panel). Quantitative RT-PCR was used to confirm the efficiency of knockdown or overexpression. (B,C) A549 (B) and ANIP-973 (C) cells were exposed to culture medium containing different concentrations of gemcitabine (0, 0.5, 1, 2, 4, 8 and 16 μg/mL). Then, cell viability was measured by CCK8 assay. (D) A549 and ANIP-973 cells were exposed to culture medium containing 2 μg/mL gemcitabine. Colony formation experiments were employed to examine the clonal formation ability. (E) Western blot analysis was used to detect the expression levels of vimentin, N-cadherin and E- cadherin in A549 (control or CYTOR-overexpressing) and ANIP-973 (control or sh-CYTOR) cells. Data are shown as the mean±SD, and the experiments were performed three times in triplicate. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Cell lines and patient tissues Human LUAD cell lines (A549, ANIP-973, and NCI-H2122 cells) and a bronchial epithelial cell line (16HBE cells) were obtained from ATCC (Manassas, USA).

Techniques: In Vitro, Quantitative RT-PCR, Knockdown, Over Expression, CCK-8 Assay, Western Blot, Expressing, Control

Journal: Acta biochimica et biophysica Sinica

Article Title: lncRNA CYTOR promotes lung adenocarcinoma gemcitabine resistance and epithelial-mesenchymal transition by sponging miR-125a-5p and upregulating ANLN and RRM2.

doi: 10.3724/abbs.2023287

Figure Lengend Snippet: Figure 3. CYTOR acts as a miRNA sponge molecule and absorbs miR-125a-5p (A) Five miRNAs (hsa-miR-525-3p, hsa-miR-125b-5p, hsa-miR- 125a-5p, hsa-miR-206, and miR-138-5p) were selected by taking the intersection of the predicted CYTOR-targeting miRNAs and downregulated miRNAs in LUAD samples from TCGA database. (B) Correlation analysis of CYTOR expression with five miRNAs in LUAD samples from the TCGA database. (C) The expression level of miR-125a-5p was evaluated using TCGA paired LUAD samples. (D) Quantitative RT-PCR was used to detect miR-125a-5p expression in eight paired LUAD and adjacent tissues. (E‒G) Relationships between miR-125a-5p expression and different T stages (T1 and T2-4) (E) and N stages (N0 and N1-3) (F) were analyzed in TCGA-LUAD samples. (H-I) Overall survival (H) and disease-specific survival (I) curve analysis in TCGA-LUAD patients with low or high miR-125a-5p expression. (J) Differentially expressed miR-125a-5p in LUAD cell lines (A549, ANIP-973, and NCI-H2122) and a human bronchial epithelial cell line (16HBE) is shown. (K) CYTOR was overexpressed in A549 cells or silenced in ANIP-973 cells, and the expression level of miR-125a-5p was measured by quantitative RT-PCR. (L) Dual-luciferase reporter assays were used to detect luciferase activity in ANIP-973 cells overexpressing miR-125a-5p. (M) RNA immunoprecipitation (RIP) of AGO2 followed by quantitative RT- PCR. The amount of miR-125a-5p bound to AGO2 or IgG (control) was measured. (N) RNA pulldown assays using biotin-labelled miR-125a-5p or control probes were performed to determine the enrichment of CYTOR in the lncRNA‒miRNA complexes. Data are shown as the mean±SD, and the experiments were performed three times in triplicate. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Cell lines and patient tissues Human LUAD cell lines (A549, ANIP-973, and NCI-H2122 cells) and a bronchial epithelial cell line (16HBE cells) were obtained from ATCC (Manassas, USA).

Techniques: Expressing, Quantitative RT-PCR, Luciferase, Activity Assay, RNA Immunoprecipitation, Control

Journal: Acta biochimica et biophysica Sinica

Article Title: lncRNA CYTOR promotes lung adenocarcinoma gemcitabine resistance and epithelial-mesenchymal transition by sponging miR-125a-5p and upregulating ANLN and RRM2.

doi: 10.3724/abbs.2023287

Figure Lengend Snippet: Figure 4. miR-125a-5p binds to ANLN/RRM2 mRNAs and attenuates their expression (A) A total of 122 mRNAs were obtained by taking the intersection of the predicted miR-125a-5p-targeting genes based on online bioinformatics tools (PITA, RNA22, miRmap, microT, miRanda, PicTar, and TargetScan) and genes upregulated in LUAD. (B-C) The associations between ANLN/RRM2 and miR-125a-5p expression (B) and ANLN/RRM2 and CYTOR expression (C) were analyzed using the TCGA-LUAD dataset. (D) Quantitative RT-PCR was performed to measure the expressions of miR-125a-5p, ANLN and RRM2 in A549 cells with miR-125a-5p silencing and in ANIP-973 cells with miR-125a-5p overexpression. (E) Western blot analysis of ANLN and RRM2 expressions in A549 cells transfected with CYTOR- or miR-125a-5p-overexpressing vectors. (F–G) The binding sites and binding mutations of ANLN (ANLN WT and ANLN MUT) (F)/RRM2 (RRM2 WT and RRM2 MUT) (G) with miR-125a-5p were cloned and inserted into the luciferase reporter vector pmirGLO and transfected into ANIP-973 cells. Then, dual-luciferase reporter assays were used to detect luciferase activity in ANIP-973 cells with or without miR-125a-5p overexpression. (H) RIP assays using AGO2 antibodies or normal IgG (negative control) followed by quantitative RT-PCR were performed in ANIP-973 cells with or without miR-125a-5p overexpression. The relative enrichment of ANLN and RRM2 RNAs is shown. (I) Biotin-labelled miRNA pulldowns were conducted to verify the interaction of miR-125a-5p with ANLN and RRM2. Data are shown as the mean±SD, and the experiments were performed three times in triplicate. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Cell lines and patient tissues Human LUAD cell lines (A549, ANIP-973, and NCI-H2122 cells) and a bronchial epithelial cell line (16HBE cells) were obtained from ATCC (Manassas, USA).

Techniques: Expressing, Quantitative RT-PCR, Over Expression, Western Blot, Transfection, Binding Assay, Clone Assay, Luciferase, Plasmid Preparation, Activity Assay, Negative Control

Journal: Acta biochimica et biophysica Sinica

Article Title: lncRNA CYTOR promotes lung adenocarcinoma gemcitabine resistance and epithelial-mesenchymal transition by sponging miR-125a-5p and upregulating ANLN and RRM2.

doi: 10.3724/abbs.2023287

Figure Lengend Snippet: Figure 5. ANLN and RRM2 act as oncogenic genes in LUAD (A) The expression levels of ANLN and RRM2 were analyzed based on LUAD nonpaired (left) and paired (right) subtypes from the TCGA database. (B–E) Comparison of ANLN and RRM2 expression in LUAD patients with different T stages (T1 and T2-4) (B), N stages (N0 and N1-3) (C), M stages (M0 and M1) (D), and pathological stages (I, II and III & IV) (E) from the TCGA LUAD subtypes. (F–G) The protein expression levels of ANLN (F) and RRM2 (G) in LUAD were analyzed according to the CPTAC database. (H–I) Kaplan‒Meier survival analysis was employed based on the TCGA database. Overall survival, disease-specific survival, and progression-free survival were analyzed with respect to ANLN (H) and RRM2 (I) expression levels. (J) The receiver operating characteristic (ROC) curve of OS associated with ANLN and RRM2 expression. (K) ANIP-973 cells with ANLN (left) or RRM2 (right) overexpression were exposed to culture medium containing different concentrations of gemcitabine (0, 0.5, 1, 2, 4, 8 and 16 μg/mL). Then, cell viability was measured by CCK8 assay. Data are shown as the mean±SD, and the experiments were performed three times in triplicate. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Cell lines and patient tissues Human LUAD cell lines (A549, ANIP-973, and NCI-H2122 cells) and a bronchial epithelial cell line (16HBE cells) were obtained from ATCC (Manassas, USA).

Techniques: Expressing, Comparison, Over Expression, CCK-8 Assay

Journal: Acta biochimica et biophysica Sinica

Article Title: lncRNA CYTOR promotes lung adenocarcinoma gemcitabine resistance and epithelial-mesenchymal transition by sponging miR-125a-5p and upregulating ANLN and RRM2.

doi: 10.3724/abbs.2023287

Figure Lengend Snippet: Figure 6. The CYTOR/miR-125a-5p/ANLN and RRM2 axis promotes gemcitabine resistance of LUAD and EMT CYTOR was stably overexpressed in A549 cells. Then, cells with CYTOR overexpression (CYTOR group) or negative control cells (vector group) were subcutaneously implanted into nude mice (n=5 per group). The mice were treated with gemcitabine or PBS (mock treatment) two weeks later (14 d). (A) The volumes of xenograft tumors in nude mice was recorded every week. (B,C) The mice were sacrificed at the sixth week (42 d) and the tumors were isolated. Tumors were imaged (B) and weighed (C). (D) IHC was used to detect Ki67, PCNA, ANLN, RRM2, N-cadherin, E-cadherin and TUNEL expression in paraffin- embedded xenograft tumor sections. (E) Diagram of the molecular mechanism of CYTOR in promoting gemcitabine resistance of LUAD and EMT by sponging miR-125a-5p and upregulating ANLN/RRM2 expression. Data are shown as the mean±SD, and the experiments were performed three times in triplicate. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Cell lines and patient tissues Human LUAD cell lines (A549, ANIP-973, and NCI-H2122 cells) and a bronchial epithelial cell line (16HBE cells) were obtained from ATCC (Manassas, USA).

Techniques: Stable Transfection, Over Expression, Negative Control, Plasmid Preparation, Isolation, TUNEL Assay, Expressing

Journal: Journal of Hematology & Oncology

Article Title: LCAT3, a novel m6A-regulated long non-coding RNA, plays an oncogenic role in lung cancer via binding with FUBP1 to activate c-MYC

doi: 10.1186/s13045-021-01123-0

Figure Lengend Snippet: LCAT3 is upregulated in lung tumor tissues and associated with poor prognosis of LUAD. A Boxplot showing the relative expression of LCAT3 in lung tumors ( n = 485) and adjacent normal ( n = 56) tissues in a TCGA LUAD cohort. LCAT3 expression was quantified by FPKM in the RNA-seq data. B LCAT3 expression validated by qRT-PCR in an independent cohort of 13 paired samples of LUAD tissues and adjacent normal tissues. LCAT3 expression was normalized to the expression of β-actin. C , D Kaplan–Meier curves of overall survival ( C ) and disease-free survival ( D ) of lung cancer patients with high versus low expressions of LCAT3. E , F The protein coding potential of LCAT3 evaluated by the Coding Potential Assessment Tool (CPAT) and Coding Potential Calculator 2 (CPC2). Two verified lncRNAs, XIST and HORAIR, served as controls for the prediction. G , H Subcellular localization of LCAT3. Real-time PCR analysis confirmed the nuclear and cytoplasmic fraction of LCAT3 transcript in Calu1 and Hop62 cells; U6 and GAPDH served as positive controls for the nuclear and cytoplasmic fractions, respectively

Article Snippet: The human LUAD cell lines (A549, Calu1 and Hop62) and a human embryonic kidney cell line (HEK-293T) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

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

Journal: Journal of Hematology & Oncology

Article Title: LCAT3, a novel m6A-regulated long non-coding RNA, plays an oncogenic role in lung cancer via binding with FUBP1 to activate c-MYC

doi: 10.1186/s13045-021-01123-0

Figure Lengend Snippet: LCAT3 is upregulated by METTL3 through m6A modification. A METTL3 is upregulated in LUAD. Boxplot showing the relative expression of METTL3 in lung tumors ( n = 480) and adjacent normal tissues ( n = 56) in TCGA LUAD cohort. METTL3 expression was quantified by FPKM in the RNA-seq data. B Western blot assays to detect METTL3 knockout efficiency in A549 and Calu1 cells. C , D qRT-PCR analysis of LCAT3 expression in sg-lacZ and sg-METTL3 lung cancer cells. E MeRIP-seq of sgLacZ (top) and sgMETTL3 (bottom) A549 cells showed that depletion of METTL3 reduces the m6A modification on LCAT3. Red line represents input group while blue line represents IP group. F MeRIP-RT-qPCR confirmed the change in m6A levels in METTL3-knockout A549 cells. G , H Half-life of LCAT3 in sg-lacZ and sg-METTL3 lung cancer cells treated with 5 μg/mL Actinomycin D

Article Snippet: The human LUAD cell lines (A549, Calu1 and Hop62) and a human embryonic kidney cell line (HEK-293T) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Modification, Expressing, RNA Sequencing, Western Blot, Knock-Out, Quantitative RT-PCR

Journal: Journal of Hematology & Oncology

Article Title: LCAT3, a novel m6A-regulated long non-coding RNA, plays an oncogenic role in lung cancer via binding with FUBP1 to activate c-MYC

doi: 10.1186/s13045-021-01123-0

Figure Lengend Snippet: LCAT3 physically interacts with FUBP1. A Proteins interacting with LCAT3 were identified by pull down assay followed by mass spectrometry analysis. B Western blot analysis of FUBP1 in beads only, negative control, sense and antisense LCAT3 pull-down fractions. GAPDH served as a negative control. C . FUBP1 RIP assays in Calu1 cells. A western blot assay confirmed FUBP1 immunoprecipitation (Up); The relative fold enrichment of LCAT3 between FUBP1 and IgG RIP fractions was determined by qRT-PCR (Down). D LCAT3 sense and 208–342 nt fragments were performed with RNA pull down and western blotting assays. E The schematic structure of full-length FUBP1 proteins (FL: 1–644) and three domain-deleted mutants (MUT1: 100–644; MUT2: 1–100 and 447–644; MUT3: 1–511) of FUBP1 variants used in this study. The blue box is the inhibitory domain, the orange box is the DNA and RNA binding domain, and the purple box is the transcription domain. F RIP assays were performed using anti-FLAG antibodies in Calu1 cells that were transfected with vectors expressing the FLAG-tagged FL and the deleted mutants (MUT1-3) of FUBP1. G Relative mRNA expression of FUBP1 primary tumors ( n = 515) and in adjacent normal tissues ( n = 59) from TCGA LUAD samples. H Relative protein expression of FUBP1 in primary tumors ( n = 111) and adjacent normal tissue ( n = 111) from Clinical Proteomic Tumor Analysis Consortium (CPTAC) LUAD samples. I Kaplan–Meier survival analysis of overall survival of lung cancer patients using FUBP1 expression. The data were downloaded from the study of Takeuchi et al. (GEO accession: GSE11969; No. patients: 90)

Article Snippet: The human LUAD cell lines (A549, Calu1 and Hop62) and a human embryonic kidney cell line (HEK-293T) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Pull Down Assay, Mass Spectrometry, Western Blot, Negative Control, Immunoprecipitation, Quantitative RT-PCR, RNA Binding Assay, Transfection, Expressing