nonsmall cell lung cancer nsclc cell lines a549  (ATCC)

Journal: Frontiers in Oncology

Article Title: RNA Demethylase ALKBH5 Prevents Lung Cancer Progression by Regulating EMT and Stemness via Regulating p53

doi: 10.3389/fonc.2022.858694

Figure Lengend Snippet: Enrichment of CSCs from A549 and PC-9 cells. (A) After being cultured in a serum-free medium, spheres derived from A549 and PC-9 were imaged at an amplification of ×10. (B) By performing CCK-8 assay, the cell viability of CSCs and their parental cells from days 1 to 5 was measured. * p < 0.05 vs. the parental cell group. The mRNA (C) and protein levels (D) of stemness hallmarkers, including CD24, CD44, ALDH1, Nanog, Oct4, and Sox2 were measured by performing RT-qPCR and Western blot analysis. * p < 0.05 vs. parental cell group.

Article Snippet: Nonsmall cell lung cancer (NSCLC) cell lines A549 and PC-9 were all purchased from the American Type Culture Collection (ATCC).

Techniques: Cell Culture, Derivative Assay, Amplification, CCK-8 Assay, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Oncology

Article Title: RNA Demethylase ALKBH5 Prevents Lung Cancer Progression by Regulating EMT and Stemness via Regulating p53

doi: 10.3389/fonc.2022.858694

Figure Lengend Snippet: CSCs present a relatively lower m 6 A methylation level compared with their parental cells. (A) Global m 6 A methylation was measured in CSCs and its parental cells of A549 and PC-9. * p < 0.05 vs. parental cells. m 6 A methylation-relative gene expressions including METLL3, METLL14, YTHDF1, WTAP, FTO, and ALKBH5 were measured in mRNA (B) and protein levels (C) . (D) By employing the GEPIA database analysis tool, the relative mRNA levels of METLL3, METLL14, YTHDF1, WTAP, FTO, and ALKBH5 were compared between tumor samples and nontumor samples.

Article Snippet: Nonsmall cell lung cancer (NSCLC) cell lines A549 and PC-9 were all purchased from the American Type Culture Collection (ATCC).

Techniques: Methylation

Journal: Frontiers in Oncology

Article Title: RNA Demethylase ALKBH5 Prevents Lung Cancer Progression by Regulating EMT and Stemness via Regulating p53

doi: 10.3389/fonc.2022.858694

Figure Lengend Snippet: p53 transcriptionally regulates ALKBH5 and subsequently decreased m 6 A methylation. (A) GEPIA database analysis tool was used to compare the correlation between ALKBH5 and p53 in LUAD and LUSC. (B) In A549 and PC-9 CSCs, the efficiency of p53 knockdown or overexpression was measured by performing a Western blot analysis. (C) After modification of p53 with or without PFT-α, ALKBH5 mRNA and protein level were measured. * p < 0.05 vs. shScrambled group; # p < 0.05 vs. p53 group. (D) After modification of p53 with or without PFT-α, m 6 A methylation level was measured. * p < 0.05 vs. shScrambled group; # p < 0.05 vs. shp53 group (left panel); * p < 0.05, vs. vector group; # p < 0.05 vs. p53 group (right panel).

Article Snippet: Nonsmall cell lung cancer (NSCLC) cell lines A549 and PC-9 were all purchased from the American Type Culture Collection (ATCC).

Techniques: Methylation, Over Expression, Western Blot, Modification, Plasmid Preparation

Journal: Frontiers in Oncology

Article Title: RNA Demethylase ALKBH5 Prevents Lung Cancer Progression by Regulating EMT and Stemness via Regulating p53

doi: 10.3389/fonc.2022.858694

Figure Lengend Snippet: p53 inhibits malignancies via exerting transcriptional activity. In A549 CSCs, p53 was efficiently knockdown by transfecting shRNA targeting to p53 mRNA. In PC-9 CSCs, p53 was efficiently overexpressed by transfecting p53-coding plasmid. Cell viability was then analyzed by performing CCK-8 assay (A) . * p < 0.05 vs. shScrambled group (left panel); * p < 0.05 vs. vector group (right panel). (B) Cell cycle distribution was analyzed by performing PI staining followed by flow cytometry assay. * p < 0.05 vs. shScrambled group; # p < 0.05 vs. shp53 group (left panel); * p < 0.05 vs. vector group; # p < 0.05 vs. p53 group (right panel). (C) Cell invasion was analyzed by performing Transwell assay. * p <0.05 vs. shScrambled group; # p < 0.05, vs. shp53 group (left panel); * p < 0.05, vs. vector group; # p < 0.05 vs. p53 group (right panel). (D) Tumor formation in soft agar was performed. * p < 0.05 vs. shScrambled group; # p < 0.05 vs. shp53 group (left panel); * p < 0.05 vs. vector group; # p < 0.05 vs. p53 group (right panel).

Article Snippet: Nonsmall cell lung cancer (NSCLC) cell lines A549 and PC-9 were all purchased from the American Type Culture Collection (ATCC).

Techniques: Activity Assay, shRNA, Plasmid Preparation, CCK-8 Assay, Staining, Flow Cytometry, Transwell Assay

Journal: Frontiers in Oncology

Article Title: RNA Demethylase ALKBH5 Prevents Lung Cancer Progression by Regulating EMT and Stemness via Regulating p53

doi: 10.3389/fonc.2022.858694

Figure Lengend Snippet: p53 potentially regulates PRRX1, Sox2, and E-cadherin via ALKBH5. In A549 CSCs, p53 was efficiently knockdown by transfecting shRNA targeting p53 mRNA. In PC-9 CSCs, p53 was efficiently overexpressed by transfecting p53-coding plasmid. mRNA (A) and protein (B) of PRRX1, Sox2, and E-cadherin were measured. * p < 0.05 shScrambled group (left panel); * p < 0.05 vs. vector group; # p < 0.05 vs. p53 group (right panel).

Article Snippet: Nonsmall cell lung cancer (NSCLC) cell lines A549 and PC-9 were all purchased from the American Type Culture Collection (ATCC).

Techniques: shRNA, Plasmid Preparation

Journal: ACS sensors

Article Title: Sensitive Detection of Exosomal Proteins via a Compact Surface Plasmon Resonance Biosensor for Cancer Diagnosis

doi: 10.1021/acssensors.8b00230

Figure Lengend Snippet: (a) Representative real-time response of compact SPR biosensor upon the addition of water, PBS, A549 exosomes at concentration of 4 × 1010 exosomes/mL, and PBS washing buffer. (b) SEM images of the biochip surface before (left) and after (right) the capture of EGFR positive exosomes. (c) Exosomes derived from A549 lung cancer cells showed higher exosomal EGFR expression than those from BEAS-2B normal cells. Both A549 exosomes and BEAS-2B exosomes were applied on the biochip at concentration of 4 × 1010 exosomes/mL. (d) With anti-IgG control antibodies modified biochip, no significant nonspecific binding of A549 exosomes was observed. A549 exosomes were applied on the biochip at much higher concentration, i.e., 2 × 1011 exosomes/mL.

Article Snippet: A549 nonsmall cell lung cancer (NSCLC) cells and BEAS-2B normal human bronchial epithelial cells were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Concentration Assay, Derivative Assay, Expressing, Modification, Binding Assay

Journal: ACS sensors

Article Title: Sensitive Detection of Exosomal Proteins via a Compact Surface Plasmon Resonance Biosensor for Cancer Diagnosis

doi: 10.1021/acssensors.8b00230

Figure Lengend Snippet: Exosomal EGFR expression measured by ELISA (a) and compact SPR biosensor (b) at increasing A549 exosome concentrations from 0 to 4 × 1010 exosomes/mL (n = 3).

Article Snippet: A549 nonsmall cell lung cancer (NSCLC) cells and BEAS-2B normal human bronchial epithelial cells were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay