Journal: Cancer Research

Article Title: Epithelial to Mesenchymal Transition Is a Determinant of Sensitivity of Non–Small-Cell Lung Carcinoma Cell Lines and Xenografts to Epidermal Growth Factor Receptor Inhibition

doi: 10.1158/0008-5472.can-05-1058

Figure Lengend Snippet: Figure 3. A, NSCLC lines were grown as s.c. xenografts in nu/nu mice and dosed for 14 days with erlotinib (100 mg/kg qd) or vehicle control. Statistical evaluation of percent tumor growth inhibition by repeated measures ANOVA was done. H292, H358, and H441 were found to be sensitive to EGFR inhibition in vivo. No correlation between tumor growth rate and sensitivity to EGFR inhibition was observed. Solid bars, erlotinib dosing period. B, tumor cell lysates were evaluated for the expression of epithelial and mesenchymal proteins. Tumors were grown to a volume of f300 mm3, excised, and flash frozen in liquid nitrogen. Tumor tissues were pulverized while frozen, subjected to detergent lysis and SDS-PAGE as described, and immunoblots probed with antibodies to E-cadherin, h-catenin, Brk, fibronectin, and vimentin. Similar with in vitro results, E-cadherin expression was restricted to erlotinib-sensitive lines and vimentin to insensitive lines. Antibody to GAPDH was used as a loading control.

Article Snippet: The human NSCLC lines (H292, H358, H322, H441, A549, Calu6, H460, H1703, SW1573, Calu3, Colo699, H2122, A427, H1437, H1299, Hop92, and H23) were cultured in the appropriate American Type Culture Collection recommended supplemented media.

Techniques: Control, Inhibition, In Vivo, Expressing, Lysis, SDS Page, Western Blot, In Vitro

Journal: Pharmaceuticals

Article Title: Self-Assembled Supramolecular Nanoparticles Improve the Cytotoxic Efficacy of CK2 Inhibitor THN7

doi: 10.3390/ph11010010

Figure Lengend Snippet: Cytotoxic activity of THN7-loaded α-cyclodextrin nanoparticles on A-427 lung cancer cells. C4: C 4 H 9 ; C6: C 6 H 13 ; C8: C 8 H 17 . (+) loaded cyclodextrins; (−) empty cyclodextrins. Viability of A427 cells were determined using an MTT assay. The viability of cells treated with 1% of the appropriate solvent was set 100%. Staurosporine served as a control for the correctly performance of the MTT assay using A427 cells. In all experiments THN7 was used at a final concentration of 6 μM.

Article Snippet: The nanoparticles were tested on the human lung cancer cell line A427, which was obtained from the German Collection of Microorganisms and Cell Culture (DSMZ, Braunschweig, Germany).

Techniques: Activity Assay, MTT Assay, Solvent, Control, Concentration Assay

Journal: BMC Pulmonary Medicine

Article Title: Long non-coding RNA GATA6-AS1 upregulates GATA6 to regulate the biological behaviors of lung adenocarcinoma cells

doi: 10.1186/s12890-021-01521-7

Figure Lengend Snippet: The downregulated GATA6-AS1 suppressed the proliferation and migration, as well as induced the apoptosis of LUAD. a GATA6-AS1 expression in 483 LUAD tissues and 437 adjacent normal tissues was indicated from GEPIA database. b Expression of GATA6-AS1 in 35 LUAD tissues and non-tumor tissues was detected by RT-qPCR. c RT-qPCR revealed the expression of GATA6-AS1 in LUAD cells and human lung cells. d The overexpression efficiency of GATA6-AS1 was depicted through RT-qPCR. e The EdU assay was performed to evaluate effects of upregulated GATA6-AS1 on proliferative ability of A549 and H1975 cells. f , g The colony formation assay demonstrated cell proliferation after transfection with overexpressed GATA6-AS1. h The influence of GATA6-AS1 overexpression on the apoptosis of LUAD cells was examined by the TUNEL assay. i Flow cytometric analysis depicted the apoptotic rate of A549 and H1975 cells upon the upregulation of GATA6-AS1. j – k The wound healing assay was conducted to determine cell migration affected by overexpressed GATA6-AS1. l Transwell assay was performed to reveal invasion of LUAD cells transfected with pcDNA3.1 or pcDNA3.1-GATA6-AS1. * p < 0.05, ** p < 0.01

Article Snippet: The four LUAD cell lines (A427, A549, H1975, HCC827) and human lung cell line (BEAS-2B) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Migration, Expressing, Quantitative RT-PCR, Over Expression, EdU Assay, Colony Assay, Transfection, TUNEL Assay, Wound Healing Assay, Transwell Assay

Journal: BMC Pulmonary Medicine

Article Title: Long non-coding RNA GATA6-AS1 upregulates GATA6 to regulate the biological behaviors of lung adenocarcinoma cells

doi: 10.1186/s12890-021-01521-7

Figure Lengend Snippet: As the cognate sense transcript of GATA6-AS1, GATA6 was low-expressed in LUAD cells. a GEPIA database showed the expression of GATA6 in 483 LUAD tissue samples and 347 corresponding normal tissue samples. b Expression of GATA6 in 35 LUAD tissues and non-tumor tissues was detected by RT-qPCR. c The mRNA expression and protein expression of GATA6 in LUAD cells and normal lung cells were detected by RT-qPCR and western blot. d RT-qPCR analysis was performed to reveal the overexpression efficiency of GATA6 in A549 and H1975 cells. e Western blot analysis showed the expression of GATA6 at the protein level in A549 and H1975 cells after upregulation of GATA6. f LncLocator database predicted the subcellular location of GATA6-AS1. g The main distribution of GATA6-AS1 in A549 and H1975 cells was determined utilizing the subcellular fractionation assay. * p < 0.05, ** p < 0.01

Article Snippet: The four LUAD cell lines (A427, A549, H1975, HCC827) and human lung cell line (BEAS-2B) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Over Expression, Fractionation

Journal: BMC Pulmonary Medicine

Article Title: Long non-coding RNA GATA6-AS1 upregulates GATA6 to regulate the biological behaviors of lung adenocarcinoma cells

doi: 10.1186/s12890-021-01521-7

Figure Lengend Snippet: MiR-4530 was the downstream molecule of GATA6-AS1. a The RIP assay verified that GATA6-AS1 existed in A549 and H1975 cells. b Suitable miRNAs were selected with the help of bioinformatic tools. c , d The expressions of miR-4530 and miR-1200 in LUAD cells and normal lung cells were detected by RT-qPCR. e RT-qPCR analysis measured the overexpression efficiency of miR-4530 in A549 and H1975 cells. f GATA6 expression in A549 and H1975 cells after the transfection of miR-4530-mimics and miR-4530-mimics + pcDNA3.1/GATA6-AS1 was investigated using RT-qPCR. g MiR-4530 expression in A549 and H1975 cells transfected with pcDNA3.1/GATA6-AS1 was detected by RT-qPCR. h GATA6 expression in A549 and H1975 cells transfected with pcDNA3.1, pcDNA3.1/GATA6-AS1, or cotransfected with pcDNA3.1/GATA6-AS1 + miR-4530 mimics was detected by RT-qPCR. i The possible binding sites of miR-4530 and GATA6-AS1 or GATA6 were predicted using bioinformatic tools. j , k The dual‑luciferase reporter assay was conducted to detect the interaction of miR-4530 and GATA6-AS1 or GATA6 in A549 and H1975 cells. l The co-existence of GATA6-AS1, miR-4530 and GATA6 in RISC was validated by the RIP assay. * p < 0.05, ** p < 0.01

Article Snippet: The four LUAD cell lines (A427, A549, H1975, HCC827) and human lung cell line (BEAS-2B) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Over Expression, Expressing, Transfection, Binding Assay, Reporter Assay

Journal: BMC Pulmonary Medicine

Article Title: Long non-coding RNA GATA6-AS1 upregulates GATA6 to regulate the biological behaviors of lung adenocarcinoma cells

doi: 10.1186/s12890-021-01521-7

Figure Lengend Snippet: GATA6-AS1 mediated the proliferation, migration, invasion, apoptosis of LUAD cells by GATA6. a The RT-qPCR analysis measured the knockdown efficiency of GATA6 in A549 and H1975 cells. b , c The EdU assay analyzed the influence of GATA6 silence on proliferative rate of A549 and H1975 cells reduced by overexpressed GATA6-AS1. d The proliferative cells decreased by GATA6-AS1 upregulation was affected by depleting GATA6, as suggested by the colony formation assay. e The TUNEL assay was conducted to examine how GATA6-AS1 overexpression-suppressed cell apoptosis was neutralized after downregulated GATA6. f Flow cytometry analysis investigated the change of apoptotic rate in A549 and H1975 cells after transfected with upregulated GATA6-AS1 and then upregulated GATA6-AS1 plus silencing GATA6. g – h The effects of GATA6 knockdown on GATA6-AS1 upregulation-repressed cell migration and invasion were measured utilizing the wound healing and transwell assays. ** p < 0.01

Article Snippet: The four LUAD cell lines (A427, A549, H1975, HCC827) and human lung cell line (BEAS-2B) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Migration, Quantitative RT-PCR, Knockdown, EdU Assay, Colony Assay, TUNEL Assay, Over Expression, Flow Cytometry, Transfection

Journal: Chemistry & biology

Article Title: Cholesterol Seco sterol Aldehydes Induce Amyloidogenesis and Dysfunction of Wild Type Tumor Protein p53

doi: 10.1016/j.chembiol.2011.02.018

Figure Lengend Snippet: A Bar chart representing level of DNA binding nuclear p53 extracted from A-427 cells incubated at 37 °C for 24 h with either VEH (0.1 % ethanol), HNE, HHE, KA or ALD (each at 40 μM) after receiving a dose of γ-radiation (5000 rads). After the incubation, nuclear extracts were taken, total protein (BCA), p53 (Western blot) and functional p53 (ELISA) were measured. Data are reported as the mean ± SEM of triplicate determinations. Data was analyzed using a student two-tail t test and was considered significantly different from VEH (*) if P<0.05. B Upper Western blot analysis of p53 and p21 in nuclear protein extracts from A-427 cells before (−) and after (+) receiving γ-radiation (5000 rads). Lower Western blot analysis of p53 and p21 in nuclear protein extracts from A-427 cells. Cells incubated at 37 °C for 24 h with ALD (0-30 μM) after receiving a dose of γ-radiation (5000 rads). After the incubation, nuclear extracts were taken, total protein (BCA), p53 and p21 (Western blot) were measured. C Agarose gel analysis of p21 PCR DNA derived from p53 immunoprecipitation (IP) of A-427 genomic DNA. Lane 1, 100 bp ladder; Lane 2, A-427 genomic DNA with p53 IP (after irradiation); Lane 3, A-427 genomic DNA with p53 IP (after irradiation and incubated with VEH for 20 h); Lane 4, A-427 genomic DNA with p53 IP (after irradiation, ALD, 30 μM for 20 h).

Article Snippet: Measurement of p21 WAF transcription A-427 human lung carcinoma (ATCC# HTB-53) cells were grown in FCS (10 %) minimum essential medium (MEM) with L-glutamine (2mM), sodium bicarbonate (1.5 g/L), non-essential amino acids (0.1 mM), and sodium pyruvate (1 mM), to 80-90% confluency in 75 cm 2 flasks.

Techniques: Binding Assay, Incubation, Western Blot, Functional Assay, Enzyme-linked Immunosorbent Assay, IF-P, Agarose Gel Electrophoresis, Derivative Assay, Immunoprecipitation, Irradiation