Journal: Computational and Mathematical Methods in Medicine

Article Title: Protumor Effects of Histone H3–H4 Chaperone Antisilencing Feature 1B Gene on Lung Adenocarcinoma: In Silico and In Vitro Analyses

doi: 10.1155/2021/5005459

Figure Lengend Snippet: ASF1B promotes the proliferation of LUAD. (a, d) Colony formation assays, (b) CCK-8 assays depict that knockdown of ASF1B inhibits proliferation of LUAD cells. (c) Western blot analysis to evaluate the levels of proliferation signaling associated proteins' expression levels in LUAD.

Article Snippet: Non-neoplastic lung epithelial cell lines (HBE) and six LUAD cell lines (A549, H460, H1299, H1650, H1975, and PC9) were obtained from ATCC (American Type Culture Collection).

Techniques: CCK-8 Assay, Knockdown, Western Blot, Expressing

Journal: Computational and Mathematical Methods in Medicine

Article Title: Protumor Effects of Histone H3–H4 Chaperone Antisilencing Feature 1B Gene on Lung Adenocarcinoma: In Silico and In Vitro Analyses

doi: 10.1155/2021/5005459

Figure Lengend Snippet: ASF1B promotes proliferation of LUAD via regulating BCAR1. (a, b) Colony formation assays. (c, d) CCK8 assays indicating induced BCAR1 in ASF1B knockdown A549 and PC9 cells restored cell proliferation ability. (e) Restoration of BCAR1 upregulated CDK2/4 and CCND1 and decreased P27 in ASF1B knockdown cells.

Article Snippet: Non-neoplastic lung epithelial cell lines (HBE) and six LUAD cell lines (A549, H460, H1299, H1650, H1975, and PC9) were obtained from ATCC (American Type Culture Collection).

Techniques: Knockdown

Journal: Computational and Mathematical Methods in Medicine

Article Title: Protumor Effects of Histone H3–H4 Chaperone Antisilencing Feature 1B Gene on Lung Adenocarcinoma: In Silico and In Vitro Analyses

doi: 10.1155/2021/5005459

Figure Lengend Snippet: ASF1B promotes migration of LUAD cells by promoting BCAR1. (a, d) Wound healing. (b, c) Transwell assays depict increased migration ability after overexpressing BCAR1 in ASF1B knockdown LUAD cells. (e) Western blot results demonstrate decreased N-cadherin, Slug, Snail, Vimentin, and increased E-cadherin in ASF1B knockdown A549 and PC9 cells with BCAR1 overexpressed.

Article Snippet: Non-neoplastic lung epithelial cell lines (HBE) and six LUAD cell lines (A549, H460, H1299, H1650, H1975, and PC9) were obtained from ATCC (American Type Culture Collection).

Techniques: Migration, Knockdown, Western Blot

Journal: Computational and Mathematical Methods in Medicine

Article Title: Protumor Effects of Histone H3–H4 Chaperone Antisilencing Feature 1B Gene on Lung Adenocarcinoma: In Silico and In Vitro Analyses

doi: 10.1155/2021/5005459

Figure Lengend Snippet: Correlation of ASF1B with 64 types of noncancerous cell populations in LUAD.

Article Snippet: Non-neoplastic lung epithelial cell lines (HBE) and six LUAD cell lines (A549, H460, H1299, H1650, H1975, and PC9) were obtained from ATCC (American Type Culture Collection).

Techniques: Clinical Proteomics

Journal: International Journal of Medical Sciences

Article Title: STARD12/14 are diagnostic and prognostic biomarkers of lung adenocarcinoma associated with epigenetic regulation, immune infiltration and ferroptosis

doi: 10.7150/ijms.84566

Figure Lengend Snippet: Validation of the STARD12/14 expression in LUAD via RT-qPCR. (A) Differential expression of STARD12 in LUAD cell lines and human lung epithelial cell line. (B) Differential expression of STARD14 in LUAD cell lines and human lung epithelial cell line. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Human lung epithelial cell line (BEAS-2B Cell Article: No.CL-0496) and LUAD cell lines (NCI-H1299 Cell Article: No.CL-0165, A549 Cell Article: No.CL-0016 and PC9 Cell Article: No.CL-0298) were purchased from Procell Life Science & Technology Co. Ltd. (Wuhan, China) on December 10, 2021.

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Quantitative Proteomics

Journal: International Journal of Medical Sciences

Article Title: STARD12/14 are diagnostic and prognostic biomarkers of lung adenocarcinoma associated with epigenetic regulation, immune infiltration and ferroptosis

doi: 10.7150/ijms.84566

Figure Lengend Snippet: Protein expression level of STARD12/14 expression in LUAD via CPTAC and western blot. (A) STARD12 protein level in LUAD tissues compared with the normal. (B) STARD14 protein level in LUAD tissues compared with the normal. (C) Differential expression level of STARD12 in LUAD cell lines and human lung epithelial cell line. (D) Differential expression level of STARD14 in LUAD cell lines and human lung epithelial cell line. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Human lung epithelial cell line (BEAS-2B Cell Article: No.CL-0496) and LUAD cell lines (NCI-H1299 Cell Article: No.CL-0165, A549 Cell Article: No.CL-0016 and PC9 Cell Article: No.CL-0298) were purchased from Procell Life Science & Technology Co. Ltd. (Wuhan, China) on December 10, 2021.

Techniques: Expressing, Western Blot, Quantitative Proteomics

Journal: Open Medicine

Article Title: The function of lncRNA EMX2OS/miR-653-5p and its regulatory mechanism in lung adenocarcinoma

doi: 10.1515/med-2023-0686

Figure Lengend Snippet: Expression of EMX2OS (a) and miR-653-5p (b) in LUAD cells. The regulatory effect of miR-653-5p on the luciferase activity of EMX2OS (c) and the effect of EMX2OS on miR-653-5p expression (d). EMX2OS negatively regulated the expression of miR-653-5p in LUAD cells, and the luciferase activity of EMX2OS was negatively regulated by miR-635-5p. ** P < 0.01, *** P < 0.001, compared with normal cells or control group; ## P < 0.01 compared with the oe-EMX2OS group.

Article Snippet: LUAD cells (A549, Calu-3, PC-9, and HCC827) and normal cells (MRC5) were obtained from ATCC and cultured in the 10% FBS-containing DMEM with 0.1% penicillin and streptomycin.

Techniques: Expressing, Luciferase, Activity Assay, Control

Journal: Open Medicine

Article Title: The function of lncRNA EMX2OS/miR-653-5p and its regulatory mechanism in lung adenocarcinoma

doi: 10.1515/med-2023-0686

Figure Lengend Snippet: The function of EMX2OS and miR-653-5p in the proliferation (a) and metastasis (b) of LUAD cells. Overexpressing EMX2OS significantly suppressed cell growth and metastasis of LUAD cells, which was reversed by the overexpression of miR-653-5p. ** P < 0.01, compared with the control group; ## P < 0.01, compared with the oe-EMX2OS group.

Article Snippet: LUAD cells (A549, Calu-3, PC-9, and HCC827) and normal cells (MRC5) were obtained from ATCC and cultured in the 10% FBS-containing DMEM with 0.1% penicillin and streptomycin.

Techniques: Over Expression, Control

Journal: Cancer Cell International

Article Title: Circulating extracellular vesicle PTPRO methylation: an exploratory biomarker for minimally invasive diagnosis of early-stage lung adenocarcinoma

doi: 10.1186/s12935-025-04127-9

Figure Lengend Snippet: Methylation analysis of PTPRO in LUAD cell lines and LUAD cells derived sEVs. (A) The mRNA expression levels of PTPRO were determined by RT-qPCR in the normal lung epithelial cell line HBE and LUAD cell lines A549, NCI-H460, and PC-9. (B-C) PTPRO methylation status in HBE and the LUAD cell lines A549, NCI-H460, and PC-9 was assessed using MSP and q-MSP assays. (D) Representative TEM images of sEVs purified from the conditioned media of HBE and LUAD cell lines (A549, NCI-H460, and PC-9). Scale bar: 100 nm. (E) The concentration and size distribution of sEVs from HBE and LUAD cell lines (A549, NCI-H460, and PC-9) were evaluated using NTA. (F) Expression of sEV markers (TSG101, Alix, CD63, and Calnexin) and PTPRO in sEVs purified from HBE and LUAD cell lines (A549, NCI-H460, and PC-9) was detected by immunoblotting. (G) The mRNA expression levels of PTPRO in sEVs purified from the conditioned media of HBE and LUAD cell lines (A549, NCI-H460, and PC-9) were measured by RT-qPCR. (H-I) MSP and q-MSP analyses of purified sEVs from HBE and LUAD cell lines (A549, NCI-H460, and PC-9). H 2 O was used as a negative control. Error bars indicate SEM. *** P < 0.001 by one-way ANOVA with Dunnett's multiple comparisons test.

Article Snippet: Human LUAD cell lines (A549, PC-9, and NCI-H460) and normal human bronchial epithelial cells (HBE) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Methylation, Derivative Assay, Expressing, Quantitative RT-PCR, Purification, Concentration Assay, Western Blot, Negative Control

Journal: Heliyon

Article Title: MCF2L-AS1/miR-874-3p/STAT3 feedback loop contributes to lung adenocarcinoma cell growth and cisplatin resistance

doi: 10.1016/j.heliyon.2023.e21342

Figure Lengend Snippet: Upregulated MCF2L-AS1 promotes LUAD cell growth and cisplatin resistance. (A) The expression level of MCF2L-AS1 in human normal lung epithelial cell line (BEAS-2B) and LUAD cell lines (PC9, HCC827, A549 and NCI–H23). (B) The knockdown efficiency of sh-MCF2L-AS1#1/2 in LIAD cells. (C–E) Cell proliferation was evaluated in sh-MCF2L-AS1 transfected cells by performing CCK-8, colony formation and EdU assays. (F–G) Caspase-3 activity assay and TUNEL assay were carried out to examine cell apoptosis in LUAD MCF2L-AS1 silenced LUAD cells. (H) The migration of LUAD cells was tested with Western blot assay upon MCF2L-AS1 depletion. (I) Transwell assay was conducted to assess the invasion in LUAD cells transfected with sh-MCF2L-AS1. (J) The effect of MCF2L-AS1 knockdown on cell viability in cisplatin treated LUAD cells was measured with MTT assay. **P < 0.01.

Article Snippet: In addition, the LUAD cell line PC9 was sourced from Procell Life Science & Technology (Wuhan, China).

Techniques: Expressing, Knockdown, Transfection, CCK-8 Assay, Caspase-3 Activity Assay, TUNEL Assay, Migration, Western Blot, Transwell Assay, MTT Assay

Journal: Heliyon

Article Title: MCF2L-AS1/miR-874-3p/STAT3 feedback loop contributes to lung adenocarcinoma cell growth and cisplatin resistance

doi: 10.1016/j.heliyon.2023.e21342

Figure Lengend Snippet: MCF2L-AS1 sponges miR-874-3p in LUAD. (A–B) Subcellular fractionation and FISH assays were used to determine the distribution of MCF2L-AS1 in LUAD cells. (C) The predicted miRNAs for MCF2L-AS1 from starBase. (D) The expressions of potential miRNAs in LUAD cells. (E) MiR-874-3p expression in human normal lung epithelial cell line (BEAS-2B) and LUAD cell lines (PC9, HCC827, A549 and NCI–H23). (F) The expression level of miR-874-3p in sh-MCF2L-AS1 transfected cells. (G) RIP assay was applied to evaluate the interaction between MCF2L-AS1 and miR-874-3p. (H) The binding site between MCF2L-AS1 and miR-874-3p. (I) Transfection efficiency of miR-874-3p mimics was tested in LUAD cells. (J) Luciferase reporter assay was employed to further confirm the combination of miR-874-3p in MCF2L-AS1. **P < 0.01.

Article Snippet: In addition, the LUAD cell line PC9 was sourced from Procell Life Science & Technology (Wuhan, China).

Techniques: Fractionation, Expressing, Transfection, Binding Assay, Luciferase, Reporter Assay

Journal: Heliyon

Article Title: MCF2L-AS1/miR-874-3p/STAT3 feedback loop contributes to lung adenocarcinoma cell growth and cisplatin resistance

doi: 10.1016/j.heliyon.2023.e21342

Figure Lengend Snippet: STAT3 is a target of miR-874-3p. (A) Potential target mRNAs of miR-874-3p predicted by PITA, PicTar, miRanda, microT and miRmap. (B) Underlying downstream mRNAs that could be upregulated in LUAD cells and downregulated by miR-874-3p mimics. (C) RNA pull-down assay was utilized to examine the enrichment of STAT3, TRIB2 and SLC12A5 in miR-874-3p biotin probe. (D) The expression of STAT3 in human normal lung epithelial cell line (BEAS-2B) and LUAD cell lines (PC9, HCC827, A549 and NCI–H23). (E) The interaction between miR-874-3p together with MCF2L-AS1 and STAT3 was determined by RIP assay. (F) The potential binding site between miR-874-3p and STAT3 was predicted through starBase. (G) MiR-874-3p was verified to combine with STAT3 through luciferase reporter assay. (H–I) STAT3 mRNA and protein levels were assessed in LUAD cells transfected with miR-874-3p mimics. (J–K) qRT-PCR and Western blot assays were employed to detect the mRNA and protein levels of STAT3 in MCF2L-AS1 downregulated LUAD cells. **P < 0.01.

Article Snippet: In addition, the LUAD cell line PC9 was sourced from Procell Life Science & Technology (Wuhan, China).

Techniques: Pull Down Assay, Expressing, Binding Assay, Luciferase, Reporter Assay, Transfection, Quantitative RT-PCR, Western Blot

Journal: Heliyon

Article Title: MCF2L-AS1/miR-874-3p/STAT3 feedback loop contributes to lung adenocarcinoma cell growth and cisplatin resistance

doi: 10.1016/j.heliyon.2023.e21342

Figure Lengend Snippet: MCF2L-AS1 facilitate LUAD cell growth by targeting miR-874-3p/STAT3. (A) Expressions of miR-874-3p and STAT3 were tested by separately transfecting miR-874-3p inhibitor and sh-STAT3. (B–D) The proliferative capacity of MCF2L-AS1 silenced cells was estimated after transfecting NC inhibitor, miR-874-3p inhibitor, miR-874-3p inhibitor plus sh-STAT3. (E–F) NC inhibitor, miR-874-3p inhibitor, miR-874-3p inhibitor plus sh-STAT3 were transfected into MCF2L-AS1 downregulated cells to observe cell apoptosis. (G) Cell migration was evaluated after transfecting above appointed plasmids in sh-MCF2L-AS1 transfected cells. (H) Above appointed plasmids were transfected into MCF2L-AS1 downregulated cells to detect cell invasion. **P < 0.01.

Article Snippet: In addition, the LUAD cell line PC9 was sourced from Procell Life Science & Technology (Wuhan, China).

Techniques: Transfection, Migration

Journal: Heliyon

Article Title: MCF2L-AS1/miR-874-3p/STAT3 feedback loop contributes to lung adenocarcinoma cell growth and cisplatin resistance

doi: 10.1016/j.heliyon.2023.e21342

Figure Lengend Snippet: STAT3 binds to MCF2L-AS1 promoter. (A–B) The overexpression efficiency of STAT3 was detected in LUAD cells. (C) MCF2L-AS1 expression in STAT3 overexpressed LUAD cells. (D) The effect of STAT3 knockdown on MCF2L-AS1 expression. (E) The binding sites between STAT3 and MCF2L-AS1 promoter. (F) ChIP assay was performed to determine the interaction between STAT3 and MCF2L-AS1 promoter. (G) Luciferase reporter assay was utilized to measure the combination of STAT3 in MCF2L-AS1 promoter. **P < 0.01.

Article Snippet: In addition, the LUAD cell line PC9 was sourced from Procell Life Science & Technology (Wuhan, China).

Techniques: Over Expression, Expressing, Knockdown, Binding Assay, Luciferase, Reporter 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) 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: Computational and Mathematical Methods in Medicine

Article Title: Upregulation of NDUFAF2 in Lung Adenocarcinoma Is a Novel Independent Prognostic Biomarker

doi: 10.1155/2023/2912968

Figure Lengend Snippet: The expression of NDUFAF2 at the protein level in LUAD tissues was confirmed using clinical tissue and NSCLC cell lines. (a) Validation of the expression level of NDUFAF2 in LUAD and paired adjacent lung tissues was conducted using western blotting analysis. (b) Western blotting analysis was used to detect the protein expression level of NDUFAF2 in different NSCLC cell lines and normal lung epithelial cell line 2B. (c) IHC staining of NDUFAF2 was performed in adenocarcinoma tissue and paired adjacent normal lung tissue from typical clinical specimens. Representative images are shown. Scale bar, left, 200 μ m; right, 50 μ m.

Article Snippet: Six types of human LUAD cell lines (PC9, H358, A549, HCC827, H1299, and H1975) and pulmonary epithelial cell line (BEAS-2B) were provided by the Procell Life Science &Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing, Biomarker Discovery, Western Blot, Immunohistochemistry

Journal: Frontiers in Pharmacology

Article Title: ARID1A deficiency attenuates the response to EGFR-TKI treatment in lung adenocarcinoma

doi: 10.3389/fphar.2025.1582005

Figure Lengend Snippet: Immunohistochemical staining for ARID1A expression in EGFR-mutant LUAD tissues (50×and 200×). (A) ARID1A low expression. (B) ARID1A high expression.

Article Snippet: LUAD cell line PC-9 (Procell Life Science, Wuhan, China) was maintained in RPMI-1640 medium supplemented with 10% FBS under standard humidified conditions (37°C, 5% CO 2 ).

Techniques: Immunohistochemical staining, Staining, Expressing, Mutagenesis

Journal: Frontiers in Pharmacology

Article Title: ARID1A deficiency attenuates the response to EGFR-TKI treatment in lung adenocarcinoma

doi: 10.3389/fphar.2025.1582005

Figure Lengend Snippet: SWI/SNF subunit mutations in lung adenocarcinoma (LUAD). (A) The frequencies of EGFR and SWI/SNF subunit mutations in LUAD. (B) The frequency of SWI/SNF subunit mutations in EGFR-mutant LUAD. (C) The types of SWI/SNF subunit mutations in EGFR-mutant LUAD.

Article Snippet: LUAD cell line PC-9 (Procell Life Science, Wuhan, China) was maintained in RPMI-1640 medium supplemented with 10% FBS under standard humidified conditions (37°C, 5% CO 2 ).

Techniques: Mutagenesis

Journal: Frontiers in Pharmacology

Article Title: ARID1A deficiency attenuates the response to EGFR-TKI treatment in lung adenocarcinoma

doi: 10.3389/fphar.2025.1582005

Figure Lengend Snippet: The role of SWI/SNF subunit mutations in the prognosis of EGFR-mutant LUAD. (A-F) ARID1A, ARID1B, ARID2, ARID5B, SMARCA4, and SMARCB1.

Article Snippet: LUAD cell line PC-9 (Procell Life Science, Wuhan, China) was maintained in RPMI-1640 medium supplemented with 10% FBS under standard humidified conditions (37°C, 5% CO 2 ).

Techniques: Mutagenesis

Journal: Frontiers in Pharmacology

Article Title: ARID1A deficiency attenuates the response to EGFR-TKI treatment in lung adenocarcinoma

doi: 10.3389/fphar.2025.1582005

Figure Lengend Snippet: ARID1A mutation confers a poor prognosis for patients with EGFR-mutant LUAD. (A) The frequencies of genes exhibiting coexisting mutations with EGFR. Survival analysis for patients with the EGFR mutation, ARID1A/EGFR comutation and (B) TP53/EGFR, (C) KRAS/EGFR, (D) CDKN2A/EGFR, (E) PIK3CA/EGFR, (F) RB1/EGFR, and (G) PTEN/EGFR comutations.

Article Snippet: LUAD cell line PC-9 (Procell Life Science, Wuhan, China) was maintained in RPMI-1640 medium supplemented with 10% FBS under standard humidified conditions (37°C, 5% CO 2 ).

Techniques: Mutagenesis

Journal: Frontiers in Pharmacology

Article Title: ARID1A deficiency attenuates the response to EGFR-TKI treatment in lung adenocarcinoma

doi: 10.3389/fphar.2025.1582005

Figure Lengend Snippet: The role of ARID1A expression in the prognosis of EGFR-mutant LUAD patients receiving EGFR-TKIs as a first-line treatment after postoperative progression. (A) Comparison of PFS between patients with low ARID1A expression and patients with high ARID1A expression. (B) Forest plots for the univariate and multivariate analyses of PFS.

Article Snippet: LUAD cell line PC-9 (Procell Life Science, Wuhan, China) was maintained in RPMI-1640 medium supplemented with 10% FBS under standard humidified conditions (37°C, 5% CO 2 ).

Techniques: Expressing, Mutagenesis, Comparison

Journal: Frontiers in Pharmacology

Article Title: ARID1A deficiency attenuates the response to EGFR-TKI treatment in lung adenocarcinoma

doi: 10.3389/fphar.2025.1582005

Figure Lengend Snippet: The role of ARID1A in the prognosis of EGFR-mutant LUAD patients receiving postoperative adjuvant EGFR-TKI treatments. (A) Comparison of DFS between patients with low ARID1A expression and patients with high ARID1A expression. (B) Forest plots for univariate and multivariate analyses of DFS. (C) Nomogram for the prediction of 1-, 2- and 3-year survival. (D) Calibration curves of the nomogram.

Article Snippet: LUAD cell line PC-9 (Procell Life Science, Wuhan, China) was maintained in RPMI-1640 medium supplemented with 10% FBS under standard humidified conditions (37°C, 5% CO 2 ).

Techniques: Mutagenesis, Adjuvant, Comparison, Expressing

Journal: Cancer Biology & Therapy

Article Title: Knockdown of LINC00511 enhances radiosensitivity of lung adenocarcinoma via regulating miR-497-5p/SMAD3

doi: 10.1080/15384047.2023.2165896

Figure Lengend Snippet: Upregulated LINC00511 accelerated tumorigenesis in LUAD. (a) LINC00511 expression is up-regulated in tumor samples than in the normal samples in TCGA data by GEPIA webtool. (b-c) OS (b) and RFS (c) in LUAD. OS: overall survival; RFS: disease-free survival. (d) LINC00511 expression in LUAD cell lines. (e) Transfection of si-NC or si-LINC00511 to PC-9 and A549 cell. LINC00511 downregulations in two LUAD cell lines via si-LINC00511 treatment were realized. (f-g) Cell viability of PC-9 (f) and A549 (g) cells with or without si-LINC00511 treatment. LUAD cell viability was suppressed after LINC00511 expression being inhibited. (h) Colony formation of PC-9 and A549 cells treated with si-NC or si-LINC00511. The colony number in LINC00511-transfected LUAD cells was less than negative control group. (i) Flow cytometry results indicate an upregulation of LUAD cells apoptosis occurred when LINC00511 expression was blocked. * P < .05, ** P < .01, compared with the normal group or si-NC group.

Article Snippet: Human normal lung epithelial cell-line BEAS-2B and 4 LUAD cell lines (NCI-H1975; NCI-H441; PC-9; A549) were obtained from Shanghai iCell Bioscience Inc. BEAS-2B were seeded in DMEM medium supplemented with 10% fetal bovine serum (FBS, Gibco, Waltham, MA) and 1% Pen/Strep solution.

Techniques: Expressing, Transfection, Negative Control, Flow Cytometry

Journal: Cancer Biology & Therapy

Article Title: Knockdown of LINC00511 enhances radiosensitivity of lung adenocarcinoma via regulating miR-497-5p/SMAD3

doi: 10.1080/15384047.2023.2165896

Figure Lengend Snippet: Upregulated LINC00511 in LUAD cells after irradiation treatment. (a) 24 h later, LINC00511 expression in PC-9 and A549 cells treated with different doses of IR was detected by qRT-PCR. (b) LINC00511 expression in PC-9 and A549 cells treated with 4 Gy IR was detected every 4 hours for 24 hours by qRT-PCR. ** P < .01, compared with 0 Gy group.

Article Snippet: Human normal lung epithelial cell-line BEAS-2B and 4 LUAD cell lines (NCI-H1975; NCI-H441; PC-9; A549) were obtained from Shanghai iCell Bioscience Inc. BEAS-2B were seeded in DMEM medium supplemented with 10% fetal bovine serum (FBS, Gibco, Waltham, MA) and 1% Pen/Strep solution.

Techniques: Irradiation, Expressing, Quantitative RT-PCR

Journal: Cancer Biology & Therapy

Article Title: Knockdown of LINC00511 enhances radiosensitivity of lung adenocarcinoma via regulating miR-497-5p/SMAD3

doi: 10.1080/15384047.2023.2165896

Figure Lengend Snippet: LINC00511 knockdown potentiated radiosensitivity of LUAD cells. (a) Survival fraction of PC-9 and A549 cells exposed to gradient dose of radiation. LUAD cell survival stepped down along with the radiation dose rising. (b) Survival fraction of PC-9 and A549 cells at different LINC00511 levels under gradient IR. The decline of LUAD cell survival via radiation treatment intensified further when LINC00511 expression was blocked. (c) Cell viability of PC-9 and A549 cells under different treatments. Radiation treatment and si-LINC00511 both decreased cell viability in two cell lines. And the combination of radiation treatment and si-LINC00511 transfection further inhibited LUAD cell viability compared to the groups with single treatment. (d) Colony formation of PC-9 and A549 cells in different groups. Colony numbers in si-LINC00511 group as well as si-NC+4 Gy radiation group were both smaller than the normal control. Si-LINC00511 + 4 Gy radiation presented additional suppression in colony formation. (e) Apoptosis rate of PC-9 and A549 cells with different treatments. Based on the flow cytometry, the apoptosis was activated in si-LINC00511-transfected or radiation-treated LUAD cells. In the group treated with si-LINC00511 and 4 Gy radiation, the apoptosis activation was further promoted. (f) γ-H2AX staining of PC-9 and A549 cells under different treatments. Cells treated by si-LINC00511 exhibited enhanced DNA damage in malignant cells treated by radiation. * P < .05, ** P < .01.

Article Snippet: Human normal lung epithelial cell-line BEAS-2B and 4 LUAD cell lines (NCI-H1975; NCI-H441; PC-9; A549) were obtained from Shanghai iCell Bioscience Inc. BEAS-2B were seeded in DMEM medium supplemented with 10% fetal bovine serum (FBS, Gibco, Waltham, MA) and 1% Pen/Strep solution.

Techniques: Knockdown, Expressing, Transfection, Control, Flow Cytometry, Activation Assay, Staining

Journal: Cancer Biology & Therapy

Article Title: Knockdown of LINC00511 enhances radiosensitivity of lung adenocarcinoma via regulating miR-497-5p/SMAD3

doi: 10.1080/15384047.2023.2165896

Figure Lengend Snippet: Cytoplasmic LINC00511 was an endogenous sponge of miR-497-5p in LUAD. (a) LINC00511 information in lncATLAS. LINC00511 was predicted to locate in cytoplasm. (b) Subcellular location of LINC00511. Both in PC-9 and A549 cells, LINC00511 was identified to be located in cytoplasm. (c) Targeting relationship between LINC00511 and miR-497-5p. miR-497-5p was predicted binding to LINC00511. (d) Transfection of miR-NC or miR-497-5p to PC-9 and A549 cells. miR-497-5p expression was successfully upregulated in two LUAD cell lines after transfection. (e) Luciferase assays in PC-9 and A549. (f) LINC00511 enrichment in PC-9 and A549. Luciferase assays and miRNA array testing both proved miR-497-5p binds to LINC00511. (g) miR-497-5p expression in LUAD cell lines. (h) miR-497-5p expression in PC-9 and A549 cells treated with 4 Gy IR was detected every 4 hours for 24 hours by qRT-PCR. (i) MiR-497-5p expression in PC-9 and A549 cells transfected with si-NC or si-LINC00511. LINC00511 upregulation in LUAD cells inhibited miR-497-5p expression. ** P < .01, compared with the NC group or 0 Gy group.

Article Snippet: Human normal lung epithelial cell-line BEAS-2B and 4 LUAD cell lines (NCI-H1975; NCI-H441; PC-9; A549) were obtained from Shanghai iCell Bioscience Inc. BEAS-2B were seeded in DMEM medium supplemented with 10% fetal bovine serum (FBS, Gibco, Waltham, MA) and 1% Pen/Strep solution.

Techniques: Binding Assay, Transfection, Expressing, Luciferase, Quantitative RT-PCR

Journal: Cancer Biology & Therapy

Article Title: Knockdown of LINC00511 enhances radiosensitivity of lung adenocarcinoma via regulating miR-497-5p/SMAD3

doi: 10.1080/15384047.2023.2165896

Figure Lengend Snippet: miR-497-5p overexpression promoted the radiosensitivity of LUAD cells. (a) Cell viability of PC-9 and A549 cells under different treatments. Radiation treatment and miR-497-5p both decreased cell viability in two cell lines. The combination of radiation treatment and miR-497-5p transfection further inhibited PC-9 and A549 cells viability compared to the groups with single treatment. (b-c) Colony formation of PC-9 and A549 cells in different groups. Colony numbers in miR-497-5p or 4 Gy radiation treated LUAD cells were both lower than the normal control. And the miR-497-5p+4 Gy radiation presented additional suppression in colony formation. (d-e) Apoptosis rate of PC-9 and A549 cells with different treatments. Flow cytometry results reveal that the apoptosis was activated in miR-497-5p group or 4 Gy radiation group. In miR-497-5p+4 Gy radiation group, apoptosis activation was further enhanced. * P < .05, ** P < .01.

Article Snippet: Human normal lung epithelial cell-line BEAS-2B and 4 LUAD cell lines (NCI-H1975; NCI-H441; PC-9; A549) were obtained from Shanghai iCell Bioscience Inc. BEAS-2B were seeded in DMEM medium supplemented with 10% fetal bovine serum (FBS, Gibco, Waltham, MA) and 1% Pen/Strep solution.

Techniques: Over Expression, Transfection, Control, Flow Cytometry, Activation Assay

Journal: Cancer Biology & Therapy

Article Title: Knockdown of LINC00511 enhances radiosensitivity of lung adenocarcinoma via regulating miR-497-5p/SMAD3

doi: 10.1080/15384047.2023.2165896

Figure Lengend Snippet: LINC00511 regulated SMAD3 expression via miR-497-5p. (a) Targeting relationship between miR-497-5p and SMAD3. (b) Luciferase assays in PC-9 and A549 cells. SMAD3 binding to miR-497-5p was confirmed in LUAD cells. (c) SMAD3 expression in LUAD cell lines. (d) SMAD3 expression in PC-9 and A549 cells treated with 4 Gy IR was detected every 4 hours for 24 hours by qRT-PCR. (e) 24 hours later, SMAD3 expression in PC-9 and A549 cells treated with 4 Gy IR was detected by western blot. (f-g) 48 hours later, the mRNA (f) and protein (g) level of SMAD3 in PC-9 and A549 cells transfected with si-LINC00511 was detected by qRT-PCR and western blot. (h-i) 48 hours later, the mRNA (h) and protein (i) level of SMAD3 in PC-9 and A549 cells transfected with miR-497-5p mimics was detected by qRT-PCR and western blot. * P < .05, ** P < .01, compared with the NC group.

Article Snippet: Human normal lung epithelial cell-line BEAS-2B and 4 LUAD cell lines (NCI-H1975; NCI-H441; PC-9; A549) were obtained from Shanghai iCell Bioscience Inc. BEAS-2B were seeded in DMEM medium supplemented with 10% fetal bovine serum (FBS, Gibco, Waltham, MA) and 1% Pen/Strep solution.

Techniques: Expressing, Luciferase, Binding Assay, Quantitative RT-PCR, Western Blot, Transfection

Journal: Cancer Biology & Therapy

Article Title: Knockdown of LINC00511 enhances radiosensitivity of lung adenocarcinoma via regulating miR-497-5p/SMAD3

doi: 10.1080/15384047.2023.2165896

Figure Lengend Snippet: SMAD3 suppression enhanced the radiosensitivity of LUAD cells. (a) SMAD3 mRNA expression in PC-9 and A549 cells treated with si-NC or si-SMAD3. (b) Western blot of SMAD3 in cells with or without si-SMAD3 treatment. (c) PC-9 and A549 cells’ survival fraction at different SMAD3 levels under gradient radiation treatment. The decline of LUAD cell survival via radiation dose rising was expanded after SMAD3 downregulation. (d-e) Apoptosis rate of PC-9 and A549 cells under different treatments. In accordance with flow cytometry results, apoptosis was activated in si-SMAD3-transfected or 4 Gy radiation-treated LUAD cells. In the group treated with the combination of si-SMAD3 and 4 Gy radiation, cell apoptosis was further promoted. ** P < .01, compared with the NC group.

Article Snippet: Human normal lung epithelial cell-line BEAS-2B and 4 LUAD cell lines (NCI-H1975; NCI-H441; PC-9; A549) were obtained from Shanghai iCell Bioscience Inc. BEAS-2B were seeded in DMEM medium supplemented with 10% fetal bovine serum (FBS, Gibco, Waltham, MA) and 1% Pen/Strep solution.

Techniques: Expressing, Western Blot, Flow Cytometry, Transfection

Journal: Cancer Biology & Therapy

Article Title: Knockdown of LINC00511 enhances radiosensitivity of lung adenocarcinoma via regulating miR-497-5p/SMAD3

doi: 10.1080/15384047.2023.2165896

Figure Lengend Snippet: LINC00511 inhibition activated miR-497-5p and mediately downregulated SMAD3, which further promoted radiosensitivity in LUAD cells. (a) SMAD3 mRNA expression in PC-9 and A549 cells of different groups. (b) SMAD3 protein expression in PC-9 and A549 cells with different treatments. Compared to the NC, si-LINC00511 transfection induced a sharp decline in SMAD3 expression, which was reversed with additional anti-miR-497-5p or SMAD3 treatment. (c) Cell viability of 4 Gy radiation-treated PC-9 and A549 cells with different treatments. Downregulated LINC00511 decreased LUAD cell viability, and along with the prolonged radiation time the decreasing was more significant, which was reversed with miR-497-5p inhibition or SMAD3 activation. (d) Colony formation of 4 Gy radiation-treated PC-9 and A549 cells with different treatments. LINC00511 downregulation suppressed colony formation of IR-treated LUAD cells while additional miR-497-5p inhibition or SMAD3 activation reactivated it. (e) Apoptosis rate of 4 Gy radiation-treated PC-9 and A549 cells in different groups. 4 Gy radiation-triggered apoptosis was further promoted via si-LINC00511 transfection, which was repressed with additional anti-miR-497-5p or SMAD3 treatment. ** P < .01, compared with the NC group.

Article Snippet: Human normal lung epithelial cell-line BEAS-2B and 4 LUAD cell lines (NCI-H1975; NCI-H441; PC-9; A549) were obtained from Shanghai iCell Bioscience Inc. BEAS-2B were seeded in DMEM medium supplemented with 10% fetal bovine serum (FBS, Gibco, Waltham, MA) and 1% Pen/Strep solution.

Techniques: Inhibition, Expressing, Transfection, Activation Assay