Journal: PLoS ONE

Article Title: Long Non Coding RNAs (lncRNAs) Are Dysregulated in Malignant Pleural Mesothelioma (MPM)

doi: 10.1371/journal.pone.0070940

Figure Lengend Snippet: Levels of lncRNA expression were normalised to 18S and relative expression levels compared to the average level in the control samples for MPM tissues and MeT-5A for cell lines using the 2 −ΔΔCq method. (a) Unsupervised cluster analysis of the top 44 lncRNAs found to be differentially expressed between MeT-5A and MPM (H226, H28, MSTO, MM05) cell lines using NCode Long Noncoding RNA microarrays. All cell lines were profiled in duplicate. Red = regions over-expressed, Blue = regions under-expressed. (b) Nine candidate lncRNAs were technically validated in MPM cell lines using RT-qPCR. For RT-qPCR, lncRNA expression levels were normalised to 18S and are expressed relative to MeT-5A. (c) NR_003548 and BX648695 were significantly elevated in MPM tissues compared to benign pleura. Turkey box plots have median values represented by the line within the boxes, and the 25 th and 75 th percentiles represented by the upper and lower lines of the box. (d) 7 candidate lncRNAs were biologically validated in an extended panel of 10MPM cell lines. All candidates demonstrated consistent up-regulation of expression. MPM – Malignant Pleural Mesothelioma, lncRNA – long noncoding RNA, Ctrl – Benign Pleura, * statistically significant at P<0.05 (two-tailed t-test).

Article Snippet: Human mesothelioma cell lines H28, H226, H2052, H2452 and MSTO obtained from the American Type Cell Culture repository (ATCC, Rockville, USA), MM05 (kindly provided by the UQ Thoracic Research Centre, The Prince Charles Hospital, Brisbane ) VMC6, VMC6/52A, VMC20, VMC40, VMC23 (kindly provided by Walter Berger, Institute of Cancer Research and Walter Klepetko, Division of Thoracic Surgery, Medical University of Vienna, Austria , ) were all grown in RPMI with 10% fetal bovine serum (FBS) at 37°C with 5% CO 2 .

Techniques: Expressing, Control, Quantitative RT-PCR, Two Tailed Test

Journal: Cancer Immunology, Immunotherapy

Article Title: Lipoteichoic acids from Staphylococcus aureus stimulate proliferation of human non-small-cell lung cancer cells in vitro

doi: 10.1007/s00262-017-1980-4

Figure Lengend Snippet: (a) Basal expression of TLR-2 mRNA in A549 and H226 cells ( n = 4), (b) regulation of TLR-2 mRNA expression upon stimulation with LTA in A549 and H226 cells, n = 4

Article Snippet: The human lung adenocarcinoma cell line A549 (ATCC-CCL-185) as well as the human lung squamous carcinoma cell line H226 were obtained from the American Type Culture Collection (Rockville, MD, USA) and cultured at 37 °C in a humidified atmosphere (95% air, 5% CO 2 ).

Techniques: Expressing

Journal: Cancer Immunology, Immunotherapy

Article Title: Lipoteichoic acids from Staphylococcus aureus stimulate proliferation of human non-small-cell lung cancer cells in vitro

doi: 10.1007/s00262-017-1980-4

Figure Lengend Snippet: Time-dependent increase in H226 proliferation and MTS activity by LTA. H226 cells were incubated with various concentrations of LTA from S. aureus or sham-incubation was performed (control). H226 proliferation was assessed by automatic cell counting ( a ) and metabolic activity was quantified by MTS assay ( b ). The horizontal dotted line indicates the baseline proliferation ( a ) and MTS activity ( b ) of sham-incubated cells which was set to 100%. All data are expressed as percentage of baseline proliferation. Means ± SEM of at least four independent experiments are given. Values marked with an asterisk differ significantly from controls ( p < 0.05)

Article Snippet: The human lung adenocarcinoma cell line A549 (ATCC-CCL-185) as well as the human lung squamous carcinoma cell line H226 were obtained from the American Type Culture Collection (Rockville, MD, USA) and cultured at 37 °C in a humidified atmosphere (95% air, 5% CO 2 ).

Techniques: Activity Assay, Incubation, Control, Cell Counting, MTS Assay

Journal: Nature Communications

Article Title: c ircTP63 functions as a ceRNA to promote lung squamous cell carcinoma progression by upregulating FOXM1

doi: 10.1038/s41467-019-11162-4

Figure Lengend Snippet: Characterization of circTP63 in LUSC cells. a Genomic loci of circTP63 gene. circTP63 is produced at the TP63 gene (NM_003722.4) locus containing exons 10–11. The back-splice junction of circTP63 was identified by Sanger sequencing. b PCR analysis for circTP63 and its linear isoform TP63 in cDNA and genomic DNA (gDNA). c Northern blot analysis showed the size and abundance of circTP63 in one paired sample of LUSC tumorous tissue and corresponding adjacent nontumorous tissues. M: marker. d qRT-PCR for the abundance of circTP63 and TP63 in H1703 cells treated with Actinomycin D at the indicated time point. e Levels of circTP63 in the nuclear and cytoplasmic fractions of SW900 and H1703 cells. The error bars ( d , e ) represent s.d. ( n = 3)

Article Snippet: All of human LUSC cells (NCI-H2170, NCI-H1703, NCI-H226, NCI-H520, SW900, SK-MES-1, BEAS-2B, and HFL-1) were purchased from the American Type Culture Collection (ATCC) and were tested negative for mycoplasma contamination.

Techniques: Produced, Sequencing, Northern Blot, Marker, Quantitative RT-PCR

Journal: Nature Communications

Article Title: c ircTP63 functions as a ceRNA to promote lung squamous cell carcinoma progression by upregulating FOXM1

doi: 10.1038/s41467-019-11162-4

Figure Lengend Snippet: c ircTP63 promotes cell proliferation and tumor growth both in vitro and in vivo. a Expression levels of circTP63 and TP63 in SW900 and H1703 cells treated with circTP63 siRNA. b Expression levels of circTP63 and TP63 in H226 and H2170 cells after transduction with circTP63 lentivirus. c and d Cell proliferation analysis of LUSC cells with silencing or stably overexpressing circTP63 . e and f Cell cycle analysis of LUSC cells with silencing or stably overexpressing circTP63 . g The volume and weight of subcutaneous xenograft tumors of H2170 cells isolated from nude mice. h The volume and weight of subcutaneous xenograft tumors of H1703 cells isolated from nude mice; center line: median of data; Bounds of box: the second quartile to the third quartile; Whisker: minimum value to maximum value. The error bars a – h represent s.d. (in a – f , n = 3; in g and h , n = 6). * p < 0.05; ** p < 0.01; *** p < 0.001, two-tailed t -test. Source data are provided as a Source Data file

Article Snippet: All of human LUSC cells (NCI-H2170, NCI-H1703, NCI-H226, NCI-H520, SW900, SK-MES-1, BEAS-2B, and HFL-1) were purchased from the American Type Culture Collection (ATCC) and were tested negative for mycoplasma contamination.

Techniques: In Vitro, In Vivo, Expressing, Transduction, Stable Transfection, Cell Cycle Assay, Isolation, Whisker Assay, Two Tailed Test

Journal: Nature Communications

Article Title: c ircTP63 functions as a ceRNA to promote lung squamous cell carcinoma progression by upregulating FOXM1

doi: 10.1038/s41467-019-11162-4

Figure Lengend Snippet: c ircTP63 contributes to cell proliferation through targeting FOXM1. a Co-expression network of circTP63 with associated 25 mRNAs. A round node represents a protein-coding gene and the arrow node represents circTP63 ( hsa_circ_0068515 ). Lines between two nodes indicate interactions between two genes. Color represents the number of lines. b A heatmap shows mRNA levels of these 25 co-expression genes in the five paired LUSC samples of SBC Human ceRNA Array analysis. c Expression analysis for FOXM1 in additional 35 paired LUSC samples. d Correlation analysis revealed positive correlation between the levels of circTP63 and FOXM1 mRNA in the tumorous tissues of the 35 LUSC patients. ΔCt values were normalized according to β-actin . e The levels of circTP63 expression and FOXM1 protein in eight paired LUSC samples. f The mRNA and protein levels of FOXM1 in the LUSC cells with knockdown or overexpression of circTP63 . g Cell proliferation assay for H226 and H2170 cells with circTP63 overexpression and FOXM1 knockdown. h Cell proliferation assay for H1703 cells with circTP63 knockdown and FOXM1 overexpression. The error bars c , e – h represent s.d. (in c , n = 35; in e – h , n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001, two-tailed t -test. Source data are provided as a Source Data file

Article Snippet: All of human LUSC cells (NCI-H2170, NCI-H1703, NCI-H226, NCI-H520, SW900, SK-MES-1, BEAS-2B, and HFL-1) were purchased from the American Type Culture Collection (ATCC) and were tested negative for mycoplasma contamination.

Techniques: Expressing, Knockdown, Over Expression, Proliferation Assay, Two Tailed Test

Journal: Nature Communications

Article Title: c ircTP63 functions as a ceRNA to promote lung squamous cell carcinoma progression by upregulating FOXM1

doi: 10.1038/s41467-019-11162-4

Figure Lengend Snippet: CENPA and CENPB are regulated by cicrTP63 through FOXM1. a The mRNA and protein levels of cell cycle-related genes in H226 and H2170 cells with circTP63 overexpression. b Expression changes of CENPA , CENPB , and CCNB1 after knockdown of FOXM1 in H2170 cells with circTP63 overexpression. c Cell proliferation assay for H226 and H2170 cells with circTP63 overexpression and joint knockdown of CENPA and CENPB . d Hypothesis diagram illustrates function and mechanism of circTP63 in LUSC progress. The error bars a – c represent s.d. ( n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001, two-tailed t -test. Source data are provided as a Source Data file

Article Snippet: All of human LUSC cells (NCI-H2170, NCI-H1703, NCI-H226, NCI-H520, SW900, SK-MES-1, BEAS-2B, and HFL-1) were purchased from the American Type Culture Collection (ATCC) and were tested negative for mycoplasma contamination.

Techniques: Over Expression, Expressing, Knockdown, Proliferation Assay, Two Tailed Test