luad cell lines a549  (ATCC)

Journal: Frontiers in Oncology

Article Title: TSPAN32 as a biomarker associated with radiotherapy and immune microenvironment remodeling in lung adenocarcinoma

doi: 10.3389/fonc.2026.1724489

Figure Lengend Snippet: Overexpression of TSPAN32 inhibits proliferation and migration of lung cancer cells and enhances their radiosensitivity. (A) 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showing TSPAN32 knockdown (si-TSPAN32) promotes while overexpression (oe-TSPAN32) suppresses A549 and H1299 cell proliferation. (B, C) Wound healing assay revealing the effect of TSPAN32 on cell migration. B: Representative images of wound closure in A549 and H1299 cells at 24 h after wounding; C: Quantification. (D, E) Clonogenic assay: surviving fraction after irradiation in TSPAN32-overexpressing vs control cells. D: Representative images; E: Quantification. (F) Apoptosis in A549 and H1299 cells after TSPAN32 modulation and 4 Gy X−ray irradiation. (G-I) Subcutaneous A549 xenografts: TSPAN32 overexpression markedly slowed tumor growth. G: Representative tumors; H: Final tumor weights; I: Growth curves over time. pcDNA3.1+ serves as the control group; oe-TSPAN32 refers to the overexpression group, abbreviated as OE; si-TSPAN32 represents the TSPAN32-targeted interference group (small interfering RNA), abbreviated as SI. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The normal lung cell BEAS-2B, LUAD cell lines A549 and H1299, and the Human Embryonic Kidney 293T cells (HEK-293T) were obtained from American Type Culture Collection (ATCC).

Techniques: Over Expression, Migration, MTT Assay, Knockdown, Wound Healing Assay, Clonogenic Assay, Irradiation, Control, Small Interfering RNA

Journal: Frontiers in Oncology

Article Title: TSPAN32 as a biomarker associated with radiotherapy and immune microenvironment remodeling in lung adenocarcinoma

doi: 10.3389/fonc.2026.1724489

Figure Lengend Snippet: The regulatory mechanism of TSPAN32 in LUAD through stabilization of PTEN. (A) KEGG enrichment analysis of differentially expressed genes. (B) Gene set enrichment analysis (GSVA) of high- and low-risk groups based on risk scores. Green: low-risk group; Orange: high-risk group. (C) GSEA analysis showing pathways enriched in the low-risk group, with significance threshold set at FDR q < 0.05. (D) Co-immunoprecipitation data for the TSPAN32-PTEN interaction. (E) Real-time quantitative PCR (RT-qPCR) analysis of the expression levels of TSPAN32, PTEN, AKT, FOXO1, PD-L1 , and VEGF genes in A549 and H1299 cells after TSPAN32 overexpression. (F) RT-qPCR analysis of the expression levels of TSPAN32, PTEN, AKT, FOXO1, PD-L1 , and VEGF genes in subcutaneous tumor tissues of nude mice. (G) Western blotting (WB) analysis of PTEN and AKT protein expression levels in A549 and H1299 cells after TSPAN32 overexpression. (H) WB analysis of PTEN, AKT, and mTOR protein expression levels in A549 and H1299 cells after ionizing radiation (4 Gy). (I) Schematic diagram illustrating the role of the TSPAN32-PTEN signaling pathway in regulating the sensitivity of LUAD.

Article Snippet: The normal lung cell BEAS-2B, LUAD cell lines A549 and H1299, and the Human Embryonic Kidney 293T cells (HEK-293T) were obtained from American Type Culture Collection (ATCC).

Techniques: Immunoprecipitation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Over Expression, Western Blot

Journal: Translational Cancer Research

Article Title: Comprehensive analysis identifies PKMYT1 as an oncogene and potential prognostic and immunological biomarker in lung adenocarcinoma

doi: 10.21037/tcr-2025-1640

Figure Lengend Snippet: PKMYT1 promotes proliferation in LUAD. (A) Western blot analysis confirmed that PKMYT1 protein expression was significantly higher in LUAD tumor tissues compared to matched adjacent normal tissues (P<0.05). (B) qPCR analysis showed elevated PKMYT1 mRNA expression in a cohort of LUAD tissues compared to normal tissues (P<0.01). (C) PKMYT1 expression was upregulated in the LUAD cell lines A549 and H1299 compared to the normal bronchial epithelial cell line B2B (P<0.01). (D-I,M,N) siRNA-mediated knockdown of PKMYT1 significantly reduced both its mRNA and protein levels in A549 and H1299 cells (P<0.05). (J,O) PKMYT1 knockdown inhibited the proliferation of A549 and H1299 cells, as measured by CCK-8 assays (P<0.05). (K,L,P,Q) EdU incorporation assays further confirmed that PKMYT1 knockdown markedly decreased the proliferation rates of both cell lines (P<0.05). Statistical significance is denoted as follows: *, P<0.05; **, P<0.01; ***, P<0.001. CCK-8, cell counting kit-8; EdU, 5-ethynyl-2'-deoxyuridine; LUAD, lung adenocarcinoma; NC, negative control; qPCR, quantitative polymerase chain reaction.

Article Snippet: The human bronchial epithelial cell line BEAS-2B (Baidi Biotech, Hangzhou, China; Catalog: C5382), the LUAD cell line A549 (Procell, Wuhan, China; Catalog: CL-0016), and the non-small cell lung cancer cell line NCI-H1299 (Procell, Catalog: CL-0165) were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, California, USA; Catalog: C11995-500BT), supplemented with 10% fetal bovine serum (FBS, Baidi Biotech, Catalog: F801-500) and 1% penicillin-streptomycin solution (Beyotime, Shanghai, China; Catalog: C0222).

Techniques: Western Blot, Expressing, Knockdown, CCK-8 Assay, Cell Counting, Negative Control, Real-time Polymerase Chain Reaction

Journal: Translational Cancer Research

Article Title: Comprehensive analysis identifies PKMYT1 as an oncogene and potential prognostic and immunological biomarker in lung adenocarcinoma

doi: 10.21037/tcr-2025-1640

Figure Lengend Snippet: PKMYT1 knockdown suppresses migration and invasion in LUAD cells. (A,B) Transwell assays show that PKMYT1 knockdown significantly reduced migration and invasion in A549 cells (P<0.01) (stained with crystal violet). (E,F) Similar effects were observed in H1299 cells (P<0.001). (C,D,G,H) Wound healing assays confirm decreased migration rates in A549 and H1299 cells after PKMYT1 knockdown (P<0.01). (I-L) Western blot analysis reveals increased E-cadherin and decreased N-cadherin and Vimentin levels, indicating reversal of EMT (P<0.01). Statistical significance is denoted as follows: *, P<0.05; **, P<0.01; ***, P<0.001. EMT, epithelial-mesenchymal transition; LUAD, lung adenocarcinoma; NC, negative control.

Article Snippet: The human bronchial epithelial cell line BEAS-2B (Baidi Biotech, Hangzhou, China; Catalog: C5382), the LUAD cell line A549 (Procell, Wuhan, China; Catalog: CL-0016), and the non-small cell lung cancer cell line NCI-H1299 (Procell, Catalog: CL-0165) were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, California, USA; Catalog: C11995-500BT), supplemented with 10% fetal bovine serum (FBS, Baidi Biotech, Catalog: F801-500) and 1% penicillin-streptomycin solution (Beyotime, Shanghai, China; Catalog: C0222).

Techniques: Knockdown, Migration, Staining, Western Blot, Negative Control

Journal: Translational Cancer Research

Article Title: Comprehensive analysis identifies PKMYT1 as an oncogene and potential prognostic and immunological biomarker in lung adenocarcinoma

doi: 10.21037/tcr-2025-1640

Figure Lengend Snippet: PKMYT1 knockdown promotes apoptosis in LUAD cells. (A-H) Flow cytometry analysis shows significantly increased apoptosis in A549 and H1299 cells following PKMYT1 knockdown (P<0.01). (I-L) Western blot analysis reveals upregulation of pro-apoptotic Bax and downregulation of anti-apoptotic Bcl-2 in both cell lines (P<0.01 for A549, P<0.05 for H1299). Statistical significance is denoted as follows: *, P<0.05; **, P<0.01; ***, P<0.001. LUAD, lung adenocarcinoma; NC, negative control.

Article Snippet: The human bronchial epithelial cell line BEAS-2B (Baidi Biotech, Hangzhou, China; Catalog: C5382), the LUAD cell line A549 (Procell, Wuhan, China; Catalog: CL-0016), and the non-small cell lung cancer cell line NCI-H1299 (Procell, Catalog: CL-0165) were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, California, USA; Catalog: C11995-500BT), supplemented with 10% fetal bovine serum (FBS, Baidi Biotech, Catalog: F801-500) and 1% penicillin-streptomycin solution (Beyotime, Shanghai, China; Catalog: C0222).

Techniques: Knockdown, Flow Cytometry, Western Blot, Negative Control