Journal: BMC Immunology

Article Title: Comparative efficacy of glucocorticoid receptor agonists on Th2 cell function and attenuation by progesterone

doi: 10.1186/s12865-020-00383-8

Figure Lengend Snippet: A comparison of prednisolone- and dexamethasone-mediated reduction of type-2 cytokine mRNA levels in primary Th2 cells. Fold differences in mRNA level from corticosteroid (CS) treated cells compared to vehicle are provided ( n = 3). Quantification of IL-13 mRNA following steroid treatment ( a ). Comparison of CS ability to suppress IL-13 ( b ). The half maximal inhibitory concentration (IC 50 ) for prednisolone and dexamethasone required to suppress IL-13 mRNA expression ( c ). Quantification of IL-5 mRNA following CS treatment ( d ). Comparison of the ability of either CS to suppress IL-5 ( e ). A comparison of the IC 50 values for prednisolone and dexamethasone required to suppress IL-5 mRNA expression ( f ). Cell counts following culture in vehicle or increasing concentration of prednisolone or dexamethasone ( f ). Data represent mean and standard error. Pred, prednisolone; Dex, dexamethasone.* p < 0.05 determined by one-way ( a , d & g ) or two-way ( b & e ) RM ANOVA or t-test ( c & f )

Article Snippet: Fig. 4 Expression of the nuclear progesterone receptor (PGR) is undetectable in a Th2 cell line (CCRF-CEM) and primary Th2 cells, but present in a breast adenocarcinoma cell line (MCF-7) used as a positive control (n = 3; a ).

Techniques: Comparison, Concentration Assay, Expressing

Journal: BMC Immunology

Article Title: Comparative efficacy of glucocorticoid receptor agonists on Th2 cell function and attenuation by progesterone

doi: 10.1186/s12865-020-00383-8

Figure Lengend Snippet: A comparison of prednisolone- and dexamethasone-induced apoptosis in a Th2 cell line (CCRF-CEM). Data are expressed as percentage of 7AAD + cells ( a ) identifying dead cells. The half maximal dose (EC 50 ) for prednisolone and dexamethasone required to induce necrosis of cells ( b ). Percentage of Annexin V + 7AAD − cells, identifying apoptotic cells ( c ). A comparison of the EC 50 values for prednisolone and dexamethasone required to reach 50% maximal induction of apoptosis in cells ( d ; n = 11). Percentage of Annexin V + 7AAD − primary Th2 cells following treatment with dexamethasone, exhibiting a similar plateau effect as CCRF-CEM at 0.5 μM dexamethasone ( e ; n = 3). Comparison of CS-induced apoptosis in CCRF-CEM as fold-difference over vehicle ( f ). Data represent mean and standard error. Pred, prednisolone; Dex, dexamethasone. * p < 0.05 determined by two-way ( a & c ), t -test ( b & e ) or one-way RM ANOVA ( g )

Article Snippet: Fig. 4 Expression of the nuclear progesterone receptor (PGR) is undetectable in a Th2 cell line (CCRF-CEM) and primary Th2 cells, but present in a breast adenocarcinoma cell line (MCF-7) used as a positive control (n = 3; a ).

Techniques: Comparison

Journal: BMC Immunology

Article Title: Comparative efficacy of glucocorticoid receptor agonists on Th2 cell function and attenuation by progesterone

doi: 10.1186/s12865-020-00383-8

Figure Lengend Snippet: Corticosteroid-induced cell death is dampened by the female sex hormone progesterone. Dexamethasone-induced cell death ( a , % 7AAD + ) and apoptosis ( b , % Annexin V + 7AAD − ) of a Th2 cell line (CCRF-CEM) in the presence or absence of progesterone (2 μM, n = 5). Head-to-head comparison of apoptosis following treatment with prednisolone ( c ) or dexamethasone ( d ) with or without progesterone ( n = 4). The half maximal response (EC 50 ) for dexamethasone and prednisolone in the presence of progesterone ( e ). Influence of progesterone on the maximal response of prednisolone- or dexamethasone-induced apoptosis ( f ). Efficacy of prednisolone vs dexamethasone to induce apoptosis in the presence of prednisolone ( g ). Data represent mean and standard error. Pred, prednisolone; Dex, dexamethasone; Prog, progesterone; Pre, pretreatment. * p < 0.05 determined by one-way ( a - e ) or two-way ( f & g ) RM ANOVA

Article Snippet: Fig. 4 Expression of the nuclear progesterone receptor (PGR) is undetectable in a Th2 cell line (CCRF-CEM) and primary Th2 cells, but present in a breast adenocarcinoma cell line (MCF-7) used as a positive control (n = 3; a ).

Techniques: Comparison

Journal: BMC Immunology

Article Title: Comparative efficacy of glucocorticoid receptor agonists on Th2 cell function and attenuation by progesterone

doi: 10.1186/s12865-020-00383-8

Figure Lengend Snippet: Expression of the nuclear progesterone receptor (PGR) is undetectable in a Th2 cell line (CCRF-CEM) and primary Th2 cells, but present in a breast adenocarcinoma cell line (MCF-7) used as a positive control (n = 3; a ). Effect of progesterone with or without dexamethasone on expression of PIBF1 (n = 3, b ) . Progesterone reduced the dexamethasone-mediated increase in FKPB5 mRNA level, but had no effect when applied alone (n = 5, c ). Data represent mean and standard error. * p < 0.05 determined by one-way ANOVA

Article Snippet: Fig. 4 Expression of the nuclear progesterone receptor (PGR) is undetectable in a Th2 cell line (CCRF-CEM) and primary Th2 cells, but present in a breast adenocarcinoma cell line (MCF-7) used as a positive control (n = 3; a ).

Techniques: Expressing, Positive Control

Journal: BMC Immunology

Article Title: Comparative efficacy of glucocorticoid receptor agonists on Th2 cell function and attenuation by progesterone

doi: 10.1186/s12865-020-00383-8

Figure Lengend Snippet: RNA-sequencing of Th2 cells

Article Snippet: Fig. 4 Expression of the nuclear progesterone receptor (PGR) is undetectable in a Th2 cell line (CCRF-CEM) and primary Th2 cells, but present in a breast adenocarcinoma cell line (MCF-7) used as a positive control (n = 3; a ).

Techniques:

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: Pharmaceutics

Article Title: Synthesis and Anticancer Activity of Novel Dual Inhibitors of Human Protein Kinases CK2 and PIM-1 †

doi: 10.3390/pharmaceutics15071991

Figure Lengend Snippet: Viability of CCRF-CEM (acute lymphoblastic leukemia), K-562 (chronic myelogenous leukemia), MCF-7 (breast carcinoma), and non-cancerous Vero cells after treatment with the tested compounds. The IC 50 values were calculated using an MTT-based assay and the following equation: Y = 100/(1 + 10^((LogIC50 − X) ∗ HillSlope)).

Article Snippet: An acute lymphoblastic leukemia ALL cell line (named CCRF-CEM) was purchased from the European Collection of Authenticated Cell Cultures (ECACC), whereas MCF-7 (hormone-dependent breast adenocarcinoma), K-562 (human chronic myelogenous leukemia), and Vero cells ( Cercopithecus aethiops kidney) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: MTT 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: 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