Journal: The Kaohsiung Journal of Medical Sciences

Article Title: N6‐methyladenosine ‐mediated LINC01087 promotes lung adenocarcinoma progression by regulating miR ‐514a‐3p to upregulate centrosome protein 55

doi: 10.1002/kjm2.12879

Figure Lengend Snippet: The effects of LINC01087 knockdown on lung adenocarcinoma (LUAD) cell proliferation, migration, invasion, and epithelial–mesenchymal transition (EMT). (A) LINC01087 expression in LUAD tissues and adjacent normal tissues was predicted using the starBase database. (B) Real‐time quantitative polymerase chain reaction was used to determine LINC01087 expression levels in LUAD cell lines (H1975, A549, PC‐9, Calu‐3, and H358 cells) and human normal bronchial epithelial cells (BEAS‐2B). H1975 and A549 cells were transfected with sh‐NC or sh‐LINC01087. (C) Cell viability was determined using the cell counting kit‐8 assay. (D) Apoptosis was determined by Annexin V‐PI staining via using flow cytometry. (E, F) Cell migration and invasion were assessed by the transwell assay. (G) Protein levels of EMT‐related proteins (E‐cadherin and vimentin) and Ras homolog gene family, member A (RhoA)/Rho‐associated protein kinase1 (ROCK1) signaling pathway markers (GTP‐RhoA, total‐RhoA, and ROCK1) were determined by Western blot. Data are presented as means ± standard deviation (SD). N = 3/group. All data were obtained from three independent replicates. GAPDH, Glyceraldehyde 3‐phosphate dehydrogenase. * p <0.05; ** p <0.01; *** p <0.001.

Article Snippet: The LUAD cell line PC‐9 was purchased from iCell (Shanghai, China).

Techniques: Knockdown, Migration, Expressing, Real-time Polymerase Chain Reaction, Transfection, Cell Counting, Staining, Flow Cytometry, Transwell Assay, Western Blot, Standard Deviation

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: N6‐methyladenosine ‐mediated LINC01087 promotes lung adenocarcinoma progression by regulating miR ‐514a‐3p to upregulate centrosome protein 55

doi: 10.1002/kjm2.12879

Figure Lengend Snippet: The effects of RNA‐binding motif protein 15 (RBM15) on the m 6 A modification level and stability of LINC01087. (A) Total m 6 A modification levels in lung adenocarcinoma (LUAD) cells and human bronchial epithelial cells were determined using the methylated RNA binding protein immunoprecipitation (Me‐RIP) assay. (B) The binding relationship between RBM15 and LINC01087 was determined using the RNA immunoprecipitation (RIP) assay. RBM15 antibody was used for RIP assay to isolate RBM15‐bonded RNAs, followed by real‐time quantitative polymerase chain reaction (RT‐qPCR) using LINC01087 specific primers. (C) Levels of m 6 A modification in LINC01087 after RBM15 knockdown in LUAD cells were determined with the MeRIP assay. (D) m 6 A modification sites in LINC01087 were predicted using the SRAMP database. (E) Schematic diagram showing m 6 A modification site and synonymous mutation in LINC01087. (F) The expression level of LINC01087 in LUAD cells cotransfected with sh‐RBM15 and LINC01087‐WT/LINC01087‐MUT was determined by RT‐qPCR. (A–C) Mut, adenine residues substituted by cytosine. (G) Levels of m 6 A modification in LINC01087 in LUAD cells cotransfected with sh‐RBM15 and LINC01087‐WT or LINC01087‐MUT were determined using the Me‐RIP assay. (H) LINC01087 mRNA stability in LUAD cells after RBM15 knockdown was determined with the RNA stability assay. Data are presented as means ± SD. N = 3/group. All data were obtained from three independent replicates. * p <0.05; ** p <0.01; *** p <0.001.

Article Snippet: The LUAD cell line PC‐9 was purchased from iCell (Shanghai, China).

Techniques: RNA Binding Assay, Modification, Methylation, Immunoprecipitation, Binding Assay, RNA Immunoprecipitation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Knockdown, Mutagenesis, Expressing, Stability Assay

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: N6‐methyladenosine ‐mediated LINC01087 promotes lung adenocarcinoma progression by regulating miR ‐514a‐3p to upregulate centrosome protein 55

doi: 10.1002/kjm2.12879

Figure Lengend Snippet: The target binding relationship between LINC01087 and miR‐514a‐3p in lung adenocarcinoma (LUAD) cells. (A) The binding relationship between LINC01087 and Ago2 was determined by RNA binding protein immunoprecipitation assay. (B) The potential binding site between LINC01087 and miR‐514a‐3p was predicted using the starBase database. (C) The interaction between LINC01087 and miR‐514a‐3p was determined using the dual‐luciferase reporter assay. (D) miR‐514a‐3p expression levels following sh‐NC or sh‐LINC01087 transfection in LUAD cells were determined using real‐time quantitative polymerase chain reaction. Data are presented as means ± SD. N = 3/group. All data were obtained from three independent replicates. ** p <0.01; *** p <0.001.

Article Snippet: The LUAD cell line PC‐9 was purchased from iCell (Shanghai, China).

Techniques: Binding Assay, RNA Binding Assay, Immunoprecipitation, Luciferase, Reporter Assay, Expressing, Transfection, Real-time Polymerase Chain Reaction

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: N6‐methyladenosine ‐mediated LINC01087 promotes lung adenocarcinoma progression by regulating miR ‐514a‐3p to upregulate centrosome protein 55

doi: 10.1002/kjm2.12879

Figure Lengend Snippet: The effects of miR‐514a‐3p overexpression on lung adenocarcinoma (LUAD) cell proliferation, migration, invasion, and epithelial–mesenchymal transition. (A) Real‐time quantitative polymerase chain reaction was used to determine miR‐514a‐3p expression levels in LUAD cell lines (H1975, A549, PC‐9, Calu‐3, and H358) and human normal bronchial epithelial cells (BEAS‐2B). LUAD cells were transfected with miR‐514a‐3p mimics to overexpress miR‐514a‐3p. (B) LUAD cell viability was determined using the cell counting kit‐8 assay. (C) Apoptosis was determined by Annexin V‐PI staining with flow cytometer. (D, E) The transwell assay was used to analyze cell migration and invasion. (F) E‐cadherin, vimentin, GTP‐RhoA, total‐RhoA, and Rho‐associated protein kinase1 (ROCK1) levels in cells were determined using Western blot. Data are presented as means ± SD. N = 3/group. All data were obtained from three independent replicates. GAPDH, Glyceraldehyde 3‐phosphate dehydrogenase; RhoA, Ras homolog gene family, member A. ** p <0.01; *** p <0.001.

Article Snippet: The LUAD cell line PC‐9 was purchased from iCell (Shanghai, China).

Techniques: Over Expression, Migration, Real-time Polymerase Chain Reaction, Expressing, Transfection, Cell Counting, Staining, Flow Cytometry, Transwell Assay, Western Blot

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: N6‐methyladenosine ‐mediated LINC01087 promotes lung adenocarcinoma progression by regulating miR ‐514a‐3p to upregulate centrosome protein 55

doi: 10.1002/kjm2.12879

Figure Lengend Snippet: LINC01087 targeted miR‐514a‐3p to upregulate centrosome protein 55 (CEP55) expression in lung adenocarcinoma (LUAD) cells. (A) Common target genes binding to miR‐514a‐3p were predicted using the starBase, TargetScan, and miRTar databases. (B) Potential binding sites between miR‐514a‐3p and CEP55 were predicted using the starBase database. (C) The interaction between miR‐514a‐3p and CEP55 mRNA 3′UTR was determined using the dual‐luciferase reporter assay. (D) The interaction between miR‐514a‐3p and CEP55 mRNA 3′UTR was determined using RNA–RNA pull‐down assay with a biotinylated miR‐514a‐3p. RNA complexes bound to the miR‐514a‐3p were eluted, and the relative expression levels of CEP55 in the RNA complexes were determined using real‐time quantitative polymerase chain reaction (RT‐qPCR). (E, F) CEP55 expression levels in LUAD cells overexpressing miR‐514a‐3p were determined using RT‐qPCR and Western blot. (G) miR‐514a‐3p expression levels in LUAD cells following sh‐LINC01087 and miR‐514a‐3p inhibitor cotransfection were determined with RT‐qPCR. (H, I) CEP55 expression levels in LUAD cells after sh‐LINC01087 and miR‐514a‐3p inhibitor cotransfection were determined using RT‐qPCR and Western blot. Data are presented as means ± SD. N = 3/group. All data were obtained from three independent replicates. GAPDH, Glyceraldehyde 3‐phosphate dehydrogenase. * p <0.05; ** p <0.01; *** p <0.001.

Article Snippet: The LUAD cell line PC‐9 was purchased from iCell (Shanghai, China).

Techniques: Expressing, Binding Assay, Luciferase, Reporter Assay, Pull Down Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Cotransfection

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: N6‐methyladenosine ‐mediated LINC01087 promotes lung adenocarcinoma progression by regulating miR ‐514a‐3p to upregulate centrosome protein 55

doi: 10.1002/kjm2.12879

Figure Lengend Snippet: The inhibitory effects of miR‐514a‐3p overexpression on lung adenocarcinoma (LUAD) cell proliferation, migration, invasion, and epithelial–mesenchymal transition as well as Ras homolog gene family, member A (RhoA)/Rho‐associated protein kinase1 (ROCK1) signaling were abolished by centrosome protein 55 (CEP55) overexpression. LUAD cells were cotransfected with miR‐514a‐3p and CEP55 overexpression plasmids. (A) Cell viability was examined using cell counting kit‐8 assay. (B) Annexin V‐PI staining with flow cytometer was used to determine apoptosis. (C, D) Cell migration and invasion were determined using the transwell assay. (E) Western blot was used to determine E‐cadherin, vimentin, GTP‐RhoA, total‐RhoA, and ROCK1 levels in cells. Data are presented as means ± SD. N = 3/group. All data were obtained from three independent replicates. GAPDH, Glyceraldehyde 3‐phosphate dehydrogenase. * p <0.05; ** p <0.01; *** p <0.001.

Article Snippet: The LUAD cell line PC‐9 was purchased from iCell (Shanghai, China).

Techniques: Over Expression, Migration, Cell Counting, Staining, Flow Cytometry, Transwell Assay, Western Blot

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: N6‐methyladenosine ‐mediated LINC01087 promotes lung adenocarcinoma progression by regulating miR ‐514a‐3p to upregulate centrosome protein 55

doi: 10.1002/kjm2.12879

Figure Lengend Snippet: miR‐514a‐3p inhibition or centrosome protein 55 (CEP55) overexpression reversed the inhibitory effects of LINC01087 knockdown on lung adenocarcinoma (LUAD) cell proliferation, migration, invasion, and epithelial–mesenchymal transition. LUAD cells were cotransfected with miR‐514a‐3p knockdown/CEP55 overexpression and LINC01087 knockdown plasmids. (A, B) CEP55 expression levels in LUAD cells were determined with real‐time quantitative polymerase chain reaction and Western blot. (C) Cell viability was determined using cell counting kit‐8 assay. (D) Apoptosis was determined by flow cytometry. (E, F) The transwell assay was employed to determine cell migration and invasion. (G) E‐cadherin and vimentin levels in LUAD cells were determined using Western blot. Data are presented as means ± SD. N = 3/group. All data were obtained from three independent replicates. GAPDH, Glyceraldehyde 3‐phosphate dehydrogenase. * p <0.05; ** p <0.01; *** p <0.001.

Article Snippet: The LUAD cell line PC‐9 was purchased from iCell (Shanghai, China).

Techniques: Inhibition, Over Expression, Knockdown, Migration, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Cell Counting, Flow Cytometry, Transwell Assay

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: N6‐methyladenosine ‐mediated LINC01087 promotes lung adenocarcinoma progression by regulating miR ‐514a‐3p to upregulate centrosome protein 55

doi: 10.1002/kjm2.12879

Figure Lengend Snippet: The effects of LINC01087 knockdown on lung adenocarcinoma (LUAD) tumor growth in vivo. Nude mice were injected with LUAD cells with stable LINC01087 knockdown. (A–C) LINC01087, miR‐514a‐3p, and centrosome protein 55 (CEP55) expression levels in tumor tissues were determined by real‐time quantitative polymerase chain reaction. (D–F) Tumor tissues were collected, and tumor volume and weight were measured. (G) Ki67 and CEP55 expression levels in tumor tissues were examined using immunohistochemistry. (H) Western blot was employed to determine GTP‐RhoA, total‐RhoA, and Rho‐associated protein kinase1 (ROCK1) levels in tumor tissues. Data are presented as means ± SD. GAPDH, Glyceraldehyde 3‐phosphate dehydrogenase; RhoA, Ras homolog gene family, member A. N = 5/group. ** p <0.01; *** p <0.001.

Article Snippet: The LUAD cell line PC‐9 was purchased from iCell (Shanghai, China).

Techniques: Knockdown, In Vivo, Injection, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemistry, Western Blot

Journal: PLoS Computational Biology

Article Title: Oxygen-Driven Tumour Growth Model: A Pathology-Relevant Mathematical Approach

doi: 10.1371/journal.pcbi.1004550

Figure Lengend Snippet: (A) Growth curve fit for 4 explant models, 2 for squamous lung carcinoma and 2 for colorectal carcinoma. The xenografted cell line Calu3 shows very similar behaviour to explant models. (B) Comparison between Calu6 and Calu3 lung cell lines; a squamous lung cancer explant; and clinical tumour material analyses. The bar chart shows the proportions of microvessel density (MVD) in area (quantified from CD31), necrosis (quantified from Hematoxylin & Eosin staining) and stroma (alpha smooth muscle actin (αSMA) positive staining). (C) Images of the different tumour models stained for αSMA and counter-stained with hematoxylin.

Article Snippet: We stained tissue sections of the following tumours: Calu6 (n = 8), Calu3 (n = 7), one squamous cell lung cancer explant (n = 28) and squamous cell lung clinical cancers (n = 12) from AstraZeneca databanks.

Techniques: Comparison, Staining

Journal: PLoS Computational Biology

Article Title: Oxygen-Driven Tumour Growth Model: A Pathology-Relevant Mathematical Approach

doi: 10.1371/journal.pcbi.1004550

Figure Lengend Snippet: Parameter results ( k p and k R ′ ) for the ODM model in explant-like tumours.

Article Snippet: We stained tissue sections of the following tumours: Calu6 (n = 8), Calu3 (n = 7), one squamous cell lung cancer explant (n = 28) and squamous cell lung clinical cancers (n = 12) from AstraZeneca databanks.

Techniques: