chronic obstructive pulmonary disease (copd) bronchial epithelial cells  (Lonza)

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Mitotic Spindle Apparatus Abnormalities in Chronic Obstructive Pulmonary Disease cells: A Potential Pathway to Lung Cancer

doi: 10.1158/1940-6207.CAPR-19-0557

Figure Lengend Snippet: Effect of NNK on centrosome. Immunofluorescence images of centrosome staining in two different cell lines in interphase and metaphase stages of cell cycle. Beas-2B cells (panel A) and COPD cells (panel B) with single centrosome staining (upper panel) and abnormal number of centrosomes (>1) in response to NNK exposure (lower panel). Merged images show centrosome and DAPI staining (scale bar = 10 μm). 1 C and 1 D show two stained centrosomes at the mitotic poles (upper panel) and multiple stained centrosomes (lower panel) in both Beas-2B and COPD cells, respectively. 1 E and 1 F represent the quantitative data for abnormal centrosomes in both interphase and metaphase, respectively in both cells. The percentage of abnormal centrosomes were higher in COPD compared to Beas-2B in both interphase and metaphase (*p<0.01, ***p<0.0004, respectively). 1 G, Western blots showing an increase in PLK4 in COPD at 72 h as compared to Beas-2B which decreased after 48 hrs. 1 H, mean intensity of PLK4 increases from 48 to 72 hr. in COPD in contrast to Beas-2B, which decreased after 48 hrs.

Article Snippet: Chronic obstructive pulmonary disease cells (COPD) were purchased from Lonza® (Cat.# 195275).

Techniques: Immunofluorescence, Staining, Western Blot

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Mitotic Spindle Apparatus Abnormalities in Chronic Obstructive Pulmonary Disease cells: A Potential Pathway to Lung Cancer

doi: 10.1158/1940-6207.CAPR-19-0557

Figure Lengend Snippet: Effect of NNK on chromosome alignment. 2 A, normal metaphase with all chromosomes aligned at the metaphase plate for the vehicle treated control (upper panel) and misaligned chromosomes in response to NNK exposure in both Beas-2B and COPD cells (lower panel). 2B, Quantitative data for the misaligned chromosomes in metaphase in response to NNK exposure. Chromosome misalignment gradually increased from 48 to 72 h (****P<0.0001). 2C, confocal z sections showing vehicle and NNK treated spindles beginning at the top of the spindle and sectioning to the bottom. Numbers in the top right indicate the z distance from the top. Arrows indicate spindle poles (scale bar = 5 μm). Untreated Beas-2B and COPD cells showed two poles at z sections 1.76 and 7.76 respectively, compared to NNK treated, where two poles did not appear in the same sections rather in different z sections with first pole at 1.17 and second pole at 5.87 for Beas-2B and 6.57 and 9.55 for COPD. 2 D and 2 E, Comparison of the mean angle of rotation in vehicle treated and NNK exposed cells. The angle of rotation was significantly higher in COPD compared to Beas-2B cells (***, P < 0.001). The Δ change in angle misorientation over control is larger in the COPD treated with NNK compared to Beas-2B (p<0.0001)

Article Snippet: Chronic obstructive pulmonary disease cells (COPD) were purchased from Lonza® (Cat.# 195275).

Techniques: Control, Comparison

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Mitotic Spindle Apparatus Abnormalities in Chronic Obstructive Pulmonary Disease cells: A Potential Pathway to Lung Cancer

doi: 10.1158/1940-6207.CAPR-19-0557

Figure Lengend Snippet: Effect of NNK on mitotic genes. 3A & B shows the changes in the level of key proteins that control the mitotic apparatus. Aurora A is upregulated in response to NNK in both Beas-2B and COPD cells at 48 and 72 h in contrast to aurora B, which showed an initial increase at 48 h followed by a decline at 72 h in Beas-2B cells. Whereas aurora B showed a gradual increase from 48 h and reached a plateau at 72 h in COPD cells. PLK1 increases at 48 h and later decreased in COPD cells compared to Beas-2B, where, it exhibited a gradual increase at 48 and 72 h time points. The COPD cells showed a gradual increase of CHK1 at 48 and 72 h in response to NNK exposure, whereas not much change in CHK1 expression in Beas-2B cells at 48 and 72 h. 3C shows the persistent increase of Gamma H2AX at 48 and 72 h in COPD cells in contrast to Beas-2B, where it decreased at both time points. The level of XPC-1 gradually increased from 48 to 72 h in COPD cells in contrast to Beas-2B, where both decreased at 72 h time point. 3D centrin-2 gradually increased from 48 to 72 h in COPD cells in contrast to Beas-2B, where centrin-2 increased only at 72 h time point.

Article Snippet: Chronic obstructive pulmonary disease cells (COPD) were purchased from Lonza® (Cat.# 195275).

Techniques: Control, Expressing

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Mitotic Spindle Apparatus Abnormalities in Chronic Obstructive Pulmonary Disease cells: A Potential Pathway to Lung Cancer

doi: 10.1158/1940-6207.CAPR-19-0557

Figure Lengend Snippet: Effect of NNK on gene expression. Beas-2B and COPD cells were treated with 100nM NNK for 2 h and then washed with PBS and allowed to grow for 48, and 72 h. The treatment was conducted under low light and the experiment was repeated three times. 4A Real-time PCR analysis of NNK exposed COPD and Beas-2B shows several unique genes specific for each cell line. 4A, AURKA and AURKB increased more than 10 folds in COPD in response to NNK. In addition, MAD2L2, DCTN6 and DYNC112 showed a two fold increase. 4B, CLASP2 and HSP90B1 were upregulated and unique to Beas-2B while TUBB4A was downregulated in response to NNK exposure in Beas-2B cells. 4C, Western blot confirmed the RT-PCR data with an increase of MAD2L2 at 72 hr in COPD and a decrease of TUBB4A in Beas-2B and an increase TUBB4A in COPD. 4D, Overlap in upregulated genes between NNK exposed COPD cells and lung adenocarcinoma and squamous cell carcinoma cell lines (analysis between the different cell lines was conducted independently). AURKA, AURKB and Cyclin B1 were all upregulated in adeno carcinoma (3.5, 5.3 and 11.1, respectively) and (2.5, 11 and 6.7, respectively) for squamous in both lung cell lines and NNK treated COPD cells (12.7, 31 and 3.2, respectively).

Article Snippet: Chronic obstructive pulmonary disease cells (COPD) were purchased from Lonza® (Cat.# 195275).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Mitotic Spindle Apparatus Abnormalities in Chronic Obstructive Pulmonary Disease cells: A Potential Pathway to Lung Cancer

doi: 10.1158/1940-6207.CAPR-19-0557

Figure Lengend Snippet: A: Proposed pathway for regulation of chromosome misalignment and mitotic spindle orientation. FOXM1 as a master regulator that directly activates Aurora B and indirectly act on Aurora A through STMN1 and PLK1. In addition, it activates PLK4 and KIF20A was identified using casual network analysis. B: FOXM1 expression Beas-2B and COPD cells at 48 and 72 h; C: The western blot shows FOXM1 expression in siRNA knockdown at 48 and 72 h time points in the presence and absence of NNK in both Beas-2B and COPD cells. Knock down of FOXM1 at 72 h showed a corresponding decrease of Aurora A and B in both COPD and Beas-2B cells indicating that FOXM1 acts through aurora kinases to control spindle orientation and chromosome misalignment.

Article Snippet: Chronic obstructive pulmonary disease cells (COPD) were purchased from Lonza® (Cat.# 195275).

Techniques: Expressing, Western Blot, Knockdown, Control