Journal: PLoS ONE

Article Title: Tumor-Specific Chromosome Mis-Segregation Controls Cancer Plasticity by Maintaining Tumor Heterogeneity

doi: 10.1371/journal.pone.0080898

Figure Lengend Snippet: A , representative metaphase FISH pictures of PTEN/CEP10 and EGFR/CEP7 dual probes showing all cells carrying one copy of Chr10, an unknown chromosome with a PTEN translocation, and three cell types differing in their composition of normal and derivative Chr7 (dChr7). Arrow points to dChr7; arrowhead to normal Chr7. B , percentage of majority cells in the parental culture, derived or converted SA or NS subcultures, and the parental culture after lentiviral transductions by pTRIPZ-Vec (P-Vec), pTRIPZ-EFEMP1 with (P-E1) or after withdrawal (P-E1wd) of doxycyclin. C–D , comparison of DNA copy number variation in chromosomes 7, 8, 17, and 22 for U251 parental derived or converted NS subcultures of NS1 or SA1-NS, respectively. The Y axis is the log ratio of intensity (the ratio of test sample and normal blood) from comparative genome hybridization. Amplifications or deletions are shown by blue lines above or below the red or green areas, respectively, based on Z-score, and those with marked changes are highlighted in purple.

Article Snippet: The comparisons of U251 variants ( ) were provided by Beth Bauer (IDEXX RADIL).

Techniques: Translocation Assay, Derivative Assay, Comparison, Hybridization

Journal: PLoS ONE

Article Title: Tumor-Specific Chromosome Mis-Segregation Controls Cancer Plasticity by Maintaining Tumor Heterogeneity

doi: 10.1371/journal.pone.0080898

Figure Lengend Snippet: A , real-time qRT-PCR (right) quantification of the expression of genes associated with neural stem cell features as well as glioma cell migration and invasion, normalized to ACTB , in cells of the same cultures used for zymography, and enzyme immunometric assays (left) for quantification of VEGFA (VEGF-165) and SPP1 (Osteopontin) in conditioned medium, normalized by cell numbers. Bar and line height are mean and SD based on quantification of 3–6 sets of independent cultures. B , western blot (top) and immunocytofluorescence (bottom) of EGFR (1∶1000 from Cell Signaling) in U251 parental (P) and two clonal NS lines (NS1 and NS2). C , soft agar colony formation assay of U251 parental (P), NS1, and P-E1wd lines. D , s.c. tumorigenicity assay of cells described above, with follow-up of tumor growth as described previously . E , immunocytofluorescence analysis of NS1 and SA1-NS before and after being subjected to neural stem cell differentiation conditions described in Methods.

Article Snippet: The comparisons of U251 variants ( ) were provided by Beth Bauer (IDEXX RADIL).

Techniques: Quantitative RT-PCR, Expressing, Migration, Zymography, Western Blot, Soft Agar Assay, Tumorigenicity Assay, Cell Differentiation

Journal: PLoS ONE

Article Title: Tumor-Specific Chromosome Mis-Segregation Controls Cancer Plasticity by Maintaining Tumor Heterogeneity

doi: 10.1371/journal.pone.0080898

Figure Lengend Snippet: A , H&E images (2X) and FISH images (100X) of i.c. xenografts derived from NS1. Arrowhead , double and single arrow point to cells with 1, 2, and 3 copies of Chr7, respectively. B , comparison of cell population equilibrium in NS1 ( in vitro culture) and the derived i.c. tumors. C , fluorescence images of i.c. xenografts derived from co-implantation of RFP-labeled STIC-enriched U251-NS and GFP-labeled U251 in a 9∶1 ratio. D–E , immunofluorescence images of i.c. xenografts from co-implantation of the two lines in a 1∶99 ratio, with purple color marking BMI1 and MELK expression by RFP cells at the tumor boundary and their expression of CD133 and SPARC in vascular mimicry within the bulk tumor mass. F , confocal immunofluorescence images of i.c. xenografts derived from 100% RFP cells, with yellow color marking co-localization of RFP with CD31 or GFAP. G , Kaplan-Meier survival curves of mice after implanting a mixture of U251 parental and STIC-enriched NS subculture. Adjusted Hazard ratios (HRs) from the stratified analysis and p-value are from Cox regression analyses examining the effect of STIC percentage on survival.

Article Snippet: The comparisons of U251 variants ( ) were provided by Beth Bauer (IDEXX RADIL).

Techniques: Derivative Assay, Comparison, In Vitro, Fluorescence, Labeling, Immunofluorescence, Expressing

Journal: PLoS ONE

Article Title: Tumor-Specific Chromosome Mis-Segregation Controls Cancer Plasticity by Maintaining Tumor Heterogeneity

doi: 10.1371/journal.pone.0080898

Figure Lengend Snippet: A , schematic illustration of the working mechanism with Chr7-MS resulting in heterogeneous subpopulations and cellular phenotype inter-conversion, with STIC inhibiting growth of TMC and/or TMC stimulating growth of STIC. The Chr7 composition was shown for representative subpopulation cells in U251, based on metaphase FISH.A higher growth rate for TMC ( r 3 ) was shown compared to that for STIC ( r 2 ). Cells marked by black circles were seen in metaphase FISH analysis, suggesting their ability to grow in vitro . Cells marked by gray boxes were not seen in metaphase FISH analysis, suggesting they are unable, or have very low ability to grow in vitro . B , changes in population equilibrium from homogeneity to heterogeneity in NS1 after serial, three-day passages in SA-culture conditions, using the same cell plating density (5×10 5 /100 mm dish), with cell types determined by FISH (experimental) and then modeled using different parameters as detailed in Methods. C . changes in cell growth speed as measured or predicted by various mathematical models. D , s.c. tumorigenicity assay showing the TMC features of 3Chr7:2n,1d cells converted from STIC by increasing percentage tumor onset (a tumor size of ∼50 mm 3 ). The plot with percent uses percents computed via the Kaplan-Meier method. Log-rank test for trend with 3Chr7:2n,1d cells shows two-sided P <0.0001.

Article Snippet: The comparisons of U251 variants ( ) were provided by Beth Bauer (IDEXX RADIL).

Techniques: In Vitro, Tumorigenicity Assay

Journal: PLoS ONE

Article Title: Tumor-Specific Chromosome Mis-Segregation Controls Cancer Plasticity by Maintaining Tumor Heterogeneity

doi: 10.1371/journal.pone.0080898

Figure Lengend Snippet: U251-NS1 infected with retrovirual vectors of VEGF-165 and LacZ described previously were s.c. implanted in nude mice as described in .

Article Snippet: The comparisons of U251 variants ( ) were provided by Beth Bauer (IDEXX RADIL).

Techniques: Infection

Journal: PLoS ONE

Article Title: Cytotoxic conjugates of betulinic acid and substituted triazoles prepared by Huisgen Cycloaddition from 30-azidoderivatives

doi: 10.1371/journal.pone.0171621

Figure Lengend Snippet: Cytotoxic activities of prepared derivatives on eight tumor (including resistant) and two normal fibroblast cell lines. All other compounds prepared in this work were also tested but their activities on these 10 cell lines were higher than 50 μM which is considered inactive.

Article Snippet: The cancer cell lines were derived from T-lymphoblastic leukemia CCRF-CEM, leukemia K562 and their multiresistant counterparts expressing P-glycoprotein, MRP1 and LRP proteins (CEM-DNR, K562-TAX) [ ], solid tumors including lung (A549) and colon (HCT116, HCT116p53-/-) carcinomas, osteosarcoma cell line (U2OS), and for comparison, tests were performed on two human non-cancer fibroblast cell lines (BJ, MRC-5).

Techniques: