Journal: Journal of Cellular and Molecular Medicine

Article Title: Cul4A is an oncogene in malignant pleural mesothelioma

doi: 10.1111/j.1582-4934.2009.00971.x

Figure Lengend Snippet: Cul4A gene is amplified in mesothelioma cells. Repre-sentative pictures of FISH assay for the copy numbers of the Cul4A gene. The probe is mapped on the chromosome 13q34 (red dots). (A) Normal lymphocyte. (B) MS-1 mesothelioma cells. (C) H290 mesothelioma cells. (D) The copy numbers of the Cul4A gene of five mesothelioma cell lines, one normal mesothelial cell line (LP9) and lymphocytes were expressed as a histogram.

Article Snippet: Human mesothelioma cell lines were obtained as follows: NCI-H28, MSTO-211H from American Type Culture Collections (ATCC, Manassas, VA, USA), REN as a generous gift from Dr. Steven Albelda (University of Pennsylvania, Philadelphia, PA), NCI-H290 and MS-1 from NIH (Frederick, MD, USA).

Techniques: Amplification

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cul4A is an oncogene in malignant pleural mesothelioma

doi: 10.1111/j.1582-4934.2009.00971.x

Figure Lengend Snippet: Cul4A is overexpressed in mesothelioma cells and MPM tumors. (A) Western blot analysis results of Cul4A protein expression in normal pleural cell line (LP9) and five mesothelioma cell lines. (B) Western blot analysis results of Cul4A protein expression in 14 malignant mesothelioma tissue samples. Internal control: b-actin. Density of Cul4A bands was quantified by normalization to β-actin using LP9 as a normal control. *Denotes higher expression of Cul4A compared to normal pleural tissue. NP, normal pleural tissue; T, mesothelioma tissue.

Article Snippet: Human mesothelioma cell lines were obtained as follows: NCI-H28, MSTO-211H from American Type Culture Collections (ATCC, Manassas, VA, USA), REN as a generous gift from Dr. Steven Albelda (University of Pennsylvania, Philadelphia, PA), NCI-H290 and MS-1 from NIH (Frederick, MD, USA).

Techniques: Western Blot, Expressing, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cul4A is an oncogene in malignant pleural mesothelioma

doi: 10.1111/j.1582-4934.2009.00971.x

Figure Lengend Snippet: Down-regulation of Cul4A with shRNA in mesothelioma cells up-regulates p21 and p27 proteins. Western blot analysis of Cul4A, p53, p21, p27 and b-actin proteins after Cul4A shRNA knockdown. (A) H290 mesothelioma cells. (B) H28 mesothelioma cells. (C) MS-1 mesothelioma cells. (D) HCT116 p53 −/− colon cancer cells. EV, empty vector.

Article Snippet: Human mesothelioma cell lines were obtained as follows: NCI-H28, MSTO-211H from American Type Culture Collections (ATCC, Manassas, VA, USA), REN as a generous gift from Dr. Steven Albelda (University of Pennsylvania, Philadelphia, PA), NCI-H290 and MS-1 from NIH (Frederick, MD, USA).

Techniques: shRNA, Western Blot, Knockdown, Plasmid Preparation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cul4A is an oncogene in malignant pleural mesothelioma

doi: 10.1111/j.1582-4934.2009.00971.x

Figure Lengend Snippet: Down-regulation of Cul4A induces cell cycle arrests in GO/G1 and inhibits mesothelioma cell growth. (A) Cell cycle analysis results in H290, H28 and MS-1 mesothelioma cells after Cul4A shRNA knockdown. (B–D) Colony formation assay of H290, H28 and MS-1 mesothelioma cells. Relative colony formation in both cell lines was expressed as percentage normalized to empty vector transfected control group (bar ± S.D.) in triplet experiments. *Denotes P < 0.05. EV, empty vector.

Article Snippet: Human mesothelioma cell lines were obtained as follows: NCI-H28, MSTO-211H from American Type Culture Collections (ATCC, Manassas, VA, USA), REN as a generous gift from Dr. Steven Albelda (University of Pennsylvania, Philadelphia, PA), NCI-H290 and MS-1 from NIH (Frederick, MD, USA).

Techniques: Cell Cycle Assay, shRNA, Knockdown, Colony Assay, Plasmid Preparation, Transfection, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cul4A is an oncogene in malignant pleural mesothelioma

doi: 10.1111/j.1582-4934.2009.00971.x

Figure Lengend Snippet: G0/G1 arrest after Cul4A down-regulation is reversed by siRNA knockdown of p21 and/or p27 in mesothelioma cells. (A) Western blot analysis of p27, p21 and b-actin proteins at 72 hrs after siRNA transfection in Cul4A shRNA stably knockdown H290 cells. (B) Cell-cycle analysis at 72 hrs after siRNA transfection and (C), at 96 hrs after transfection. Percentages of cells in S, G0/G1, G2/M and sub-G1 phases were expressed as bar ± S.D. in triplet experiments. *Denotes P < 0.05. EV: empty vector. Mock: without siRNA. Ctrl: universal negative control siRNA. p21, p21 siRNA; p27,p27 siRNA; p21+p27, p21 and p27 siRNA. Cells transfected with universal negative control siRNA were used as control groups.

Article Snippet: Human mesothelioma cell lines were obtained as follows: NCI-H28, MSTO-211H from American Type Culture Collections (ATCC, Manassas, VA, USA), REN as a generous gift from Dr. Steven Albelda (University of Pennsylvania, Philadelphia, PA), NCI-H290 and MS-1 from NIH (Frederick, MD, USA).

Techniques: Knockdown, Western Blot, Transfection, shRNA, Stable Transfection, Cell Cycle Assay, Plasmid Preparation, Negative Control, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cul4A is an oncogene in malignant pleural mesothelioma

doi: 10.1111/j.1582-4934.2009.00971.x

Figure Lengend Snippet: Overexpression of ectopic Cul4A in mesothelioma cells down-regulates p21 and p27proteins and promotes cells growth. (A) Western blot analysis of Cul4A, p27, p21 and β-actin proteins in ectopic Cul4A stably transfected H28 mesothelioma cells. Density of Cul4A bands was quantified by normalization to β-actin using empty vector transfected group as a normal control. (B) Colony formation assay of H28 mesothelioma cells. Relative colony formation was expressed as percentage normalized to empty vector transfected control group (bar ± S.D.) in triplet experiments. *Denotes P < 0.05. EV, empty vector.

Article Snippet: Human mesothelioma cell lines were obtained as follows: NCI-H28, MSTO-211H from American Type Culture Collections (ATCC, Manassas, VA, USA), REN as a generous gift from Dr. Steven Albelda (University of Pennsylvania, Philadelphia, PA), NCI-H290 and MS-1 from NIH (Frederick, MD, USA).

Techniques: Over Expression, Western Blot, Stable Transfection, Transfection, Plasmid Preparation, Control, Colony Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cul4A is an oncogene in malignant pleural mesothelioma

doi: 10.1111/j.1582-4934.2009.00971.x

Figure Lengend Snippet: Cul4A promotes degradation of p21 and p27 proteins in mesothelioma cells. (A) Western blot analysis of p27, p21 and β-actin proteins in ectopic Cul4A stably transfected H28 mesothelioma cells (H28pBabeCul4A) and empty vector transfected controls (H28pBabeEV). (B) Western blot analysis of p27, p21 and β-actin proteins in Cul4A stably knockdown H28 mesothelioma cells (H28pSuperCul4A) and empty vector transfected controls (H28pSuperEV). Cells were treated with cycloheximide for the time periods indicated. Densities of p21 and p27 bands were quantified by normalization to β-actin using the 0 hr group as a normal control.

Article Snippet: Human mesothelioma cell lines were obtained as follows: NCI-H28, MSTO-211H from American Type Culture Collections (ATCC, Manassas, VA, USA), REN as a generous gift from Dr. Steven Albelda (University of Pennsylvania, Philadelphia, PA), NCI-H290 and MS-1 from NIH (Frederick, MD, USA).

Techniques: Western Blot, Stable Transfection, Transfection, Plasmid Preparation, Knockdown, Control

Journal: Lab on a chip

Article Title: Towards personalized medicine: chemosensitivity assays of patient lung cancer cell spheroids in a perfused microfluidic platform.

doi: 10.1039/c5lc00454c

Figure Lengend Snippet: Fig. 3 Experiments with MPM H2052 cell line. Percentage of microwells containing either one spheroid(dark), two spheroids (dark grey), three spheroids (grey) or four spheroids (light grey) in flat bottom microwells (A) or round bottom microwells (B). C) Comparison between percentage of microwells with a flat or round bottom microwells containing one spheroid. D) Representative images of spheroids formed in flat bottom microwells and round bottom microwells after three days in culture. Scale bar corresponds to 250 µm. For all experiments N=64. E) Images showing spheroids stained with Hoechst (blue) and Ethidium homodimer-1 (EthD1, red) indicating cell death. Scale bar corresponds to 500 µm. F) Graph representing cell mortality in spheroids after 2 days (N=16), 8 days (N=16) and 10 days (N=8) on chip. Data are represented as means ± SEM.

Article Snippet: Cell culture experiments with the mesothelioma cell line H2052 (ATCC-CRL-5915, ATCC, France) were performed according to the previously described protocol11.

Techniques: Comparison, Staining

Journal: Oncotarget

Article Title: Activation of JNK and IRE1 is critically involved in tanshinone I-induced p62 dependent autophagy in malignant pleural mesothelioma cells: implication of p62 UBA domain

doi: 10.18632/oncotarget.15336

Figure Lengend Snippet: A . Cytotoxicity of Tan I in the H28 and H2452 mesothelioma cell lines. Two mesothelioma cell lines were treated with various concentrations of Tan I (0, 5, 10, 20, and 40 μM) for 24 h. Cell viability was determined by the MTT assay. B . The inhibitory effect of Tan I on colony formation in H28, and H2452 mesothelioma cells. The colony formation assay was performed on mesothelioma cells treated with Tan I (20 μM). C . Effect of Tan I on autophagic vacuoles in H28 cells by TEM observation. H28 cells were treated with Tan I (20 μM) for 24 h, and autophagic morphology was observed by TEM. The black arrow indicates an autolysosome that contains the remnants of digested organelles. High-magnification image of the black square (bottom panel). D . Effect of Tan I on acidic autophagic vacuoles in Tan I-treated H28 cells. H28 cells were treated with Tan I for 24 h, stained with acridine orange (AO) and observed under the FLUOVIEW FV10i confocal microscopy (Olympus, Tokyo, Japan). E . Effect of Tan I (20 μM) on punctae formation of endogenous LC3II in H28 and H2452 cells by immunofluorescence. Bar scale = 10 μM, DAPI-blue, endogenous LC3-Green. F . Effect of Tan I on PARP, p62 and LC3II in H28 and H2452 mesothelioma cells. G . Effect of Tan I on the mRNA expression of p62 and LC3II in H28 cells by RT-qPCR. Total RNA was isolated from Tan 1 treated H28 cells and RT-PCR was performed as shown in Materials and Methods.

Article Snippet: The mesothelioma cell lines H28 (ATCC® CRL 5820TM), and H2452 (ATCC ® CRL-5946 TM ) were purchased from the American Type Culture Collection (ATCC).

Techniques: MTT Assay, Colony Assay, Staining, Confocal Microscopy, Immunofluorescence, Expressing, Quantitative RT-PCR, Isolation, Reverse Transcription Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Activation of JNK and IRE1 is critically involved in tanshinone I-induced p62 dependent autophagy in malignant pleural mesothelioma cells: implication of p62 UBA domain

doi: 10.18632/oncotarget.15336

Figure Lengend Snippet: A . Effect of Tan I on ER stress-related proteins such as GRP78, CHOP, IRE1 and PERK in two mesothelioma cell lines. Western blotting was performed to determine the effect of Tan I on the expression of GRP78, CHOP, PERK, ATF4, ATF6, IRE1 and β-Actin in H28 and H2452 cells. B . Time-dependent effect of Tan I on IRE1 and CHOP expression in H28 cells. C . Effect of Tan I on the phosphorylation of JNK in H28 and H2452 cells. Two mesothelioma cell lines were exposed to Tan I and Western blotting was performed to determine the effect of Tan I on JNK phosphorylation. D . Effect of the JNK inhibitor SP600125 on the expression of p62, LC3II and IRE1 in H28 cells. H28 cells were treated with Tan I (20 μM) in the presence or absence of SP600125 for 24 h. Western blotting was performed to determine the levels of LC3 conversion, and p62/SQSTM1, IRE1 and β-Actin expression.

Article Snippet: The mesothelioma cell lines H28 (ATCC® CRL 5820TM), and H2452 (ATCC ® CRL-5946 TM ) were purchased from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Expressing, Phospho-proteomics

Journal: Disease Markers

Article Title: Circulating miR-132-3p as a Candidate Diagnostic Biomarker for Malignant Mesothelioma

doi: 10.1155/2017/9280170

Figure Lengend Snippet: Characteristics of the study groups.

Article Snippet: Mesothelioma patients were recruited at the Lungenklinik Heckeshorn, HELIOS Klinikum Emil von Behring, Berlin, Germany, and in participating medical practices of the MoMar study.

Techniques: