Journal:

Article Title: Structural Analogs of Tylophora Alkaloids may not be Functional Analogs

doi: 10.1016/j.bmcl.2007.11.054

Figure Lengend Snippet: The GI 50 of tylophora alkaloids, and PBTs on the growth inhibition of HepG2, PANC-1 and CEM cells.

Article Snippet: , HepG2 GI 50 a (μM) , PANC-1 GI 50 a (μM) , CEM GI 50 a (μM).

Techniques: Inhibition

Journal: BMC Cancer

Article Title: Evaluation of bioactive sphingolipids in 4-HPR-resistant leukemia cells

doi: 10.1186/1471-2407-11-477

Figure Lengend Snippet: Acquired resistance upon long-term 4-HPR exposure . (A, left) R cell lines were continuously exposed to different 4-HPR concentrations (0.5, 3, 5, and 10 μM; cell lines named R0.5, R3, R5, and R10, respectively) and resistance verified by comparison to 48 h drug-withdrawn R cells. Data are the average ± SD of three independent experiments performed in quadruplicates, n = 12. (A, right) Parental CCRF-CEM cells and developed R cell lines were exposed to 10 μM 4-HPR and viability estimated after 48 h of drug exposure. (n = 12). ** P < 0.01; ANOVA plus Tamhane post-hoc analysis. (B) Comparative cell proliferation was determined by BrdU labeling as specified in Materials and Methods. Data are average ± SD of four independent experiments. * P < 0.05; Kruskal Wallis and Mann-Whitney test for pair-wise comparison.

Article Snippet: 4-HPR-sensitive parental leukemia cell line (CCRF-CEM) and 4-HPR-resistant cell lines (R0.5 and R10) were challenged by different anticancer drugs, such as cisplatin (0.5, 1, 2.5, or 5 μg/ml), paclitaxel (1, 5, 10, 30, or 100 nM), or adriamycin (50, 100, or 500 nM) and other insults such as UV irradiation (1, 3, or 5 mJ/cm ) or direct oxidative stress (H O 2; 10, 50, or 100 μM).

Techniques: Comparison, Labeling, MANN-WHITNEY

Journal: BMC Cancer

Article Title: Evaluation of bioactive sphingolipids in 4-HPR-resistant leukemia cells

doi: 10.1186/1471-2407-11-477

Figure Lengend Snippet: Effect of selected insults on the viability of 4-HPR-sensitive and resistant leukemia cell lines . 4-HPR-sensitive (CCRF-CEM) and derived resistant cell lines (R0.5 and R10) were exposed to several drugs or UV radiation, and cytotoxicity tested after 48 h by the XTT assay. Data are average ± SD of three independent experiments performed in quadruplicates (n = 12); * P < 0.05, ** P < 0.01; ANOVA plus Tamhane post-hoc test.

Article Snippet: 4-HPR-sensitive parental leukemia cell line (CCRF-CEM) and 4-HPR-resistant cell lines (R0.5 and R10) were challenged by different anticancer drugs, such as cisplatin (0.5, 1, 2.5, or 5 μg/ml), paclitaxel (1, 5, 10, 30, or 100 nM), or adriamycin (50, 100, or 500 nM) and other insults such as UV irradiation (1, 3, or 5 mJ/cm ) or direct oxidative stress (H O 2; 10, 50, or 100 μM).

Techniques: Derivative Assay, XTT Assay

Journal: BMC Cancer

Article Title: Evaluation of bioactive sphingolipids in 4-HPR-resistant leukemia cells

doi: 10.1186/1471-2407-11-477

Figure Lengend Snippet: Comparative endogenous SL pattern in 4-HPR-sensitive and resistant leukemia cell lines . Cell were grown with (R cell lines) or without (CCRF-CEM) 4-HPR. Endogenous TOTAL (sum of the different fatty acid chain-length species) dhCer (A, left panel) and Cer (A, right panel) levels, as well as dhSph (B), TOTAL GluCer, and LactCer (C) levels were determined by LC/MS. Data are average ± SD of four (A, B) or two (C) independent experiments. * P < 0.05; ** P < 0.01; ANOVA plus Tamhane or Bonferroni post-hoc test.

Article Snippet: 4-HPR-sensitive parental leukemia cell line (CCRF-CEM) and 4-HPR-resistant cell lines (R0.5 and R10) were challenged by different anticancer drugs, such as cisplatin (0.5, 1, 2.5, or 5 μg/ml), paclitaxel (1, 5, 10, 30, or 100 nM), or adriamycin (50, 100, or 500 nM) and other insults such as UV irradiation (1, 3, or 5 mJ/cm ) or direct oxidative stress (H O 2; 10, 50, or 100 μM).

Techniques: Liquid Chromatography with Mass Spectroscopy

Journal: BMC Cancer

Article Title: Evaluation of bioactive sphingolipids in 4-HPR-resistant leukemia cells

doi: 10.1186/1471-2407-11-477

Figure Lengend Snippet: Comparative analysis of dihydroceramide desaturase (DES) activity . Cells were incubated with the unnatural pyridinium dhCer (C12-PyrdhCer) alone or with 4-HPR (5 μM) for the selected time points and conversion into C12-PyrCer measured by LC/MS. Data are average ± SD of two independent values. ** P < 0.01; ANOVA plus Bonferroni post-hoc test.

Article Snippet: 4-HPR-sensitive parental leukemia cell line (CCRF-CEM) and 4-HPR-resistant cell lines (R0.5 and R10) were challenged by different anticancer drugs, such as cisplatin (0.5, 1, 2.5, or 5 μg/ml), paclitaxel (1, 5, 10, 30, or 100 nM), or adriamycin (50, 100, or 500 nM) and other insults such as UV irradiation (1, 3, or 5 mJ/cm ) or direct oxidative stress (H O 2; 10, 50, or 100 μM).

Techniques: Activity Assay, Incubation, Liquid Chromatography with Mass Spectroscopy

Journal: BMC Cancer

Article Title: Evaluation of bioactive sphingolipids in 4-HPR-resistant leukemia cells

doi: 10.1186/1471-2407-11-477

Figure Lengend Snippet: Comparative SL profile analysis upon drug withdrawal . R10 cells were incubated in 4-HPR-free medium for 48 h (WD = without drug) or long-term (~3 weeks) and endogenous SL profiles analyzed by LC-MS. Significance was based on R10 values. Data are average ± SD of 2-4 independent values. * P < 0.05; ** P < 0.01; ANOVA plus Tamhane or Bonferroni post-hoc test.

Article Snippet: 4-HPR-sensitive parental leukemia cell line (CCRF-CEM) and 4-HPR-resistant cell lines (R0.5 and R10) were challenged by different anticancer drugs, such as cisplatin (0.5, 1, 2.5, or 5 μg/ml), paclitaxel (1, 5, 10, 30, or 100 nM), or adriamycin (50, 100, or 500 nM) and other insults such as UV irradiation (1, 3, or 5 mJ/cm ) or direct oxidative stress (H O 2; 10, 50, or 100 μM).

Techniques: Incubation, Liquid Chromatography with Mass Spectroscopy

Journal: BMC Cancer

Article Title: Evaluation of bioactive sphingolipids in 4-HPR-resistant leukemia cells

doi: 10.1186/1471-2407-11-477

Figure Lengend Snippet: Effect of drug withdrawal on cell resistance . (A) Parental sensitive CCRF-CEM cells, resistant R10, and long-term drug withdrawn resistant (R10 longWD) cells were treated and cell death determined by annexin V-FITC/propidium iodide staining at the selected time points. Values represent the percentage of cells in each quadrant. Representative data from two independent experiments are shown. (B) A portion of each sample for SL profile analysis (Figure 5) was saved for XTT assay estimation of resistance. Briefly, R10 cells after 48 h and long-term drug withdrawal were subjected to 48 h treatment with 3 or 10 μM 4-HPR. All data were related to the corresponding untreated cells. Statistical analysis was related to R10 data. Data are average ± SD of 12 values from to three independent experiments. * P < 0.05; ** P < 0.01; ANOVA plus Tamhane post-hoc test.

Article Snippet: 4-HPR-sensitive parental leukemia cell line (CCRF-CEM) and 4-HPR-resistant cell lines (R0.5 and R10) were challenged by different anticancer drugs, such as cisplatin (0.5, 1, 2.5, or 5 μg/ml), paclitaxel (1, 5, 10, 30, or 100 nM), or adriamycin (50, 100, or 500 nM) and other insults such as UV irradiation (1, 3, or 5 mJ/cm ) or direct oxidative stress (H O 2; 10, 50, or 100 μM).

Techniques: Staining, XTT Assay

Journal: BMC Cancer

Article Title: Evaluation of bioactive sphingolipids in 4-HPR-resistant leukemia cells

doi: 10.1186/1471-2407-11-477

Figure Lengend Snippet: Analysis of 4-HPR-mediated alterations in dhSph and Sph phosphorylation . Externally added unnatural dhSph (17C-dhSph) and Sph (17C-Sph) analogues were used to estimate the secondary effects of 4-HPR-induced DES inhibition. CCRF-CEM cells were exposed to 4-HPR (10 μM) for 1 h and unnatural 17C species added for an additional 30 min incubation, as described in Methods. The effect on endogenous SLs (A) and unnatural dhSph species (B,C) analyzed by LC/MS. Comparative exogenous and endogenous SL profiles among parental (CCRF-CEM) and resistant cell lines (R0.5, R10) (D, E) were analyzed by the same methodology. Data are average ± SD of an experiment performed in duplicate.

Article Snippet: 4-HPR-sensitive parental leukemia cell line (CCRF-CEM) and 4-HPR-resistant cell lines (R0.5 and R10) were challenged by different anticancer drugs, such as cisplatin (0.5, 1, 2.5, or 5 μg/ml), paclitaxel (1, 5, 10, 30, or 100 nM), or adriamycin (50, 100, or 500 nM) and other insults such as UV irradiation (1, 3, or 5 mJ/cm ) or direct oxidative stress (H O 2; 10, 50, or 100 μM).

Techniques: Phospho-proteomics, Analogues, Inhibition, Incubation, Liquid Chromatography with Mass Spectroscopy

Journal: BMC Cancer

Article Title: Evaluation of bioactive sphingolipids in 4-HPR-resistant leukemia cells

doi: 10.1186/1471-2407-11-477

Figure Lengend Snippet: Toxicity mediated by the combination of 4-HPR with SL modulators . Parental-sensitive (CCRF-CEM) and derived-resistant (R0.5, R5, R10) cells were co-treated with 4-HPR (1 μM), DHS (5 μM; unnatural dhSph analogue), PPMP (10 μM; glucosylceramide synthase inhibitor), SAF (4 μM; another unnatural dhSph analogue), and/or SKI-II (5-20 μM) and cytotoxicity estimated after 48 h exposure using the XTT assay. Data are average ± SD of at least two independent experiments performed in quadruplicate (n ≥ 8); ** P < 0.01; ANOVA plus Tamhane post-hoc test.

Article Snippet: 4-HPR-sensitive parental leukemia cell line (CCRF-CEM) and 4-HPR-resistant cell lines (R0.5 and R10) were challenged by different anticancer drugs, such as cisplatin (0.5, 1, 2.5, or 5 μg/ml), paclitaxel (1, 5, 10, 30, or 100 nM), or adriamycin (50, 100, or 500 nM) and other insults such as UV irradiation (1, 3, or 5 mJ/cm ) or direct oxidative stress (H O 2; 10, 50, or 100 μM).

Techniques: Derivative Assay, XTT Assay

Journal: PLoS ONE

Article Title: Effects of Novel Isoform-Selective Phosphoinositide 3-Kinase Inhibitors on Natural Killer Cell Function

doi: 10.1371/journal.pone.0099486

Figure Lengend Snippet: CCRF-CEM, human acute T lymphocytic leukemia cell line was labeled with calcein AM, incubated with mouse anti-human CD4 mAb, and then co-cultured with NK cells at 10∶1 E:T ratio in the presence of indicated inhibitors for 2 h. Culture supernatants were collected and calcein fluorescence was measured and analyzed as in . The values are presented as the means ± SEM from three independent experiments. * p <0.05 and ** p <0.01, as determined by the unpaired Student's t test using Sigma Plot 10 software and as compared to the vehicle control group.

Article Snippet: CCRF-CEM, human acute T lymphocytic leukemia cell line was kindly provided by Dr. Matthew Janes (Intellikine, La Jolla, CA).

Techniques: Labeling, Incubation, Cell Culture, Fluorescence, Software