human nonsmall cell lung cancer nsclc a549  (Cell Signaling Technology Inc)

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: Journal of Ginseng Research

Article Title: Ginsenoside Rg5 overcomes chemotherapeutic multidrug resistance mediated by ABCB1 transporter: in vitro and in vivo study

doi: 10.1016/j.jgr.2018.10.007

Figure Lengend Snippet: Chemical structure and cytotoxicity of Rg5 in MDR cancer cells. (A) Chemical structure of Rg5. (B) Cytotoxicity of Rg5 alone in pairs of A2780/T or A2780 cells. (C) The cells were treated with various concentrations of docetaxel (TXT) for 48 hours in pairs of A2780/T or A2780 cells. (D) Cytotoxicity of Rg5 alone in pairs of A549/T or A549 cells. (E) The cells were treated with various concentrations of TXT for 48 hours in pairs of A549/T or A549 cells. Cell growth was determined using the SRB assay (n ≥ 3). MDR, multidrug resistance; SRB, Sulforhodamine B.

Article Snippet: Extracellular signal–regulated kinases (ERK) 1/2 and actin antibodies were purchased from Santa Cruz Biotechnology; ABCB1 (P-gp) antibodies were purchased from Calbiochem, USA; Nrf2 antibodies were purchased from Abcam, Hong Kong; other antibodies such as AKT, P-AKT, and P-ERK1/2 were purchased from Cell Signaling Technology, Inc. Human ovarian cancer cells A2780, human nonsmall cell lung cancer (NSCLC) A549 and their PTX-resistant cell line A2780/T, and A549/T were purchased from KeyGen Biotech Co., Ltd, Nanjing, China.

Techniques: Sulforhodamine B Assay

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rg5 overcomes chemotherapeutic multidrug resistance mediated by ABCB1 transporter: in vitro and in vivo study

doi: 10.1016/j.jgr.2018.10.007

Figure Lengend Snippet: Rg5 recovered sensitivity to docetaxel. Cells were treated with the indicated drugs for 48 hours and subjected to SRB assay. Rg5 reduces the IC 50 of TXT in resistant cancer cells (A2780/T) (B) and A549/T (D) but not in drug sensitive (A2780) (A) and A549 (C) . (E) Rg5 inhibited the colony formation of TXT in resistant cancer cells A2780/T in a dose-dependent manner. ## ,** p < 0.01, ### ,*** p < 0.001 vs absence of Rg5. TXT, docetaxel; SRB, Sulforhodamine B.

Article Snippet: Extracellular signal–regulated kinases (ERK) 1/2 and actin antibodies were purchased from Santa Cruz Biotechnology; ABCB1 (P-gp) antibodies were purchased from Calbiochem, USA; Nrf2 antibodies were purchased from Abcam, Hong Kong; other antibodies such as AKT, P-AKT, and P-ERK1/2 were purchased from Cell Signaling Technology, Inc. Human ovarian cancer cells A2780, human nonsmall cell lung cancer (NSCLC) A549 and their PTX-resistant cell line A2780/T, and A549/T were purchased from KeyGen Biotech Co., Ltd, Nanjing, China.

Techniques: Sulforhodamine B Assay

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rg5 overcomes chemotherapeutic multidrug resistance mediated by ABCB1 transporter: in vitro and in vivo study

doi: 10.1016/j.jgr.2018.10.007

Figure Lengend Snippet: Rg5 sensitized the chemotherapy Drug Paclitaxel, docetaxel, 5-fluorouracil, daunorubicin, and doxorubicin to ABCB1-Mediated Drug resistance A2780/T Cells.

Article Snippet: Extracellular signal–regulated kinases (ERK) 1/2 and actin antibodies were purchased from Santa Cruz Biotechnology; ABCB1 (P-gp) antibodies were purchased from Calbiochem, USA; Nrf2 antibodies were purchased from Abcam, Hong Kong; other antibodies such as AKT, P-AKT, and P-ERK1/2 were purchased from Cell Signaling Technology, Inc. Human ovarian cancer cells A2780, human nonsmall cell lung cancer (NSCLC) A549 and their PTX-resistant cell line A2780/T, and A549/T were purchased from KeyGen Biotech Co., Ltd, Nanjing, China.

Techniques:

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rg5 overcomes chemotherapeutic multidrug resistance mediated by ABCB1 transporter: in vitro and in vivo study

doi: 10.1016/j.jgr.2018.10.007

Figure Lengend Snippet: Effects of treatment of Rg5 and docetaxel on A549/T cell xenograft nude mice model. (A) The body weight was drawn to monitor the body weight and tumor volume with time after implantation . (B) Tumour growth curves were drawn to monitor the body weight and tumor volume with time after implantation. The tumor xenografts were excised. (C) The tumors were photographed on the 27th day after implantation. (D) The tumors weighted on the 27th day after implantation. The data shown are expressed as the mean ± SD for each group (n = 9 or 10), * p < 0.05, ** p < 0.01, ** p < 0.001 . CTR, control group given vehicle; SD, standard deviation; TXT, docetaxel.

Article Snippet: Extracellular signal–regulated kinases (ERK) 1/2 and actin antibodies were purchased from Santa Cruz Biotechnology; ABCB1 (P-gp) antibodies were purchased from Calbiochem, USA; Nrf2 antibodies were purchased from Abcam, Hong Kong; other antibodies such as AKT, P-AKT, and P-ERK1/2 were purchased from Cell Signaling Technology, Inc. Human ovarian cancer cells A2780, human nonsmall cell lung cancer (NSCLC) A549 and their PTX-resistant cell line A2780/T, and A549/T were purchased from KeyGen Biotech Co., Ltd, Nanjing, China.

Techniques: Standard Deviation

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