Model making a conscious decision by the bank to participate in money laundering

На мікрорівні (тобто на рівні окремого суб’єкта господарювання) легалізація коштів, отриманих злочинним шляхом, є джерелом прибутків і ризиків окремих суб’єктів. Саме тому під час дослідження процесів легалізації кримінальних доходів на макрорівні розглядається функція максимізації корисності особи, залученої до даного процесу. Згідно з традиційною теорією, власник фінансових активів розміщує їх відповідно до очікуваного прибутку та відповідних ризиків. У випадку особи, що легалізує кошти, отримані злочинним шляхом, очікуваний прибуток порівнюється з конфіденційністю (а саме з тим, що інформація про таку особу буде надана правоохоронним і податковим органам) як окремим фактором, а також імовірністю та суворістю застосування санкцій

Inhibition of Phosphatidylcholine-Specific Phospholipase C Interferes with Proliferation and Survival of Tumor Initiating Cells in Squamous Cell Carcinoma

<div><p>Purpose</p><p>The role of phosphatidylcholine-specific phospholipase C (PC-PLC), the enzyme involved in cell differentiation and proliferation, has not yet been explored in tumor initiating cells (TICs). We investigated PC-PLC expression and effects of PC-PLC inhibition in two adherent (AD) squamous carcinoma cell lines (A431 and CaSki), with different proliferative and stemness potential, and in TIC-enriched floating spheres (SPH) originated from them.</p><p>Results</p><p>Compared with immortalized non-tumoral keratinocytes (HaCaT) A431-AD cells showed 2.5-fold higher PC-PLC activity, nuclear localization of a 66-kDa PC-PLC isoform, but a similar distribution of the enzyme on plasma membrane and in cytoplasmic compartments. Compared with A431-AD, A431-SPH cells showed about 2.8-fold lower PC-PLC protein and activity levels, but similar nuclear content. Exposure of adherent cells to the PC-PLC inhibitor D609 (48h) induced a 50% reduction of cell proliferation at doses comprised between 33 and 50 μg/ml, without inducing any relevant cytotoxic effect (cell viability 95±5%). In A431-SPH and CaSki-SPH D609 induced both cytostatic and cytotoxic effects at about 20 to 30-fold lower doses (IC50 ranging between 1.2 and 1.6 μg/ml). Furthermore, D609 treatment of A431-AD and CaSki-AD cells affected the sphere-forming efficiency, which dropped in both cells, and induced down-modulation of stem-related markers mRNA levels (Oct4, Nestin, Nanog and ALDH1 in A431; Nestin and ALDH1 in CaSki cells).</p><p>Conclusions</p><p>These data suggest that the inhibition of PC-PLC activity may represent a new therapeutic approach to selectively target the most aggressive and tumor promoting sub-population of floating spheres originated from squamous cancer cells possessing different proliferative and stemness potential.</p></div

Effects of the PC-PLC inhibitor D609 on PC-PLC activity, PC-PLC protein expression and cell proliferation in HaCaT keratinocytes and in the squamous carcinoma cell line A431-AD.

<p><b>A</b>) Proliferation assays performed on cells seeded 72 hours before adding different doses of D609 at t = 0 (● = untreated cells; □ = 1.5 μg/ml, ▲ = 3 μg/ml, ▼ = 6 μg/ml, ◇ = 12.5 μg/ml, * = 25 μg/ml and ○ = 50 μg/ml) and monitored for 24h and 48h afterwards. Cell count (mean ± SD, n = 3) of live cells was obtained by Trypan blue exclusion assays and by automated cell counter, as described in the Materials and Methods section. <b>B</b>) Cell counting (mean percentage ± SD, n = 3) of either live (white columns) or dead cells (black columns) measured by Trypan blue exclusion test in the cultures used for the proliferation assays shown in panel A. <b>C</b>) Top panel: PC-PLC activity (mean ± SD, n = 3) measured by Amplex Red assay in total lysates of control (untreated = black columns) or 50 μg/ml D609-treated cells (grey columns). Statistical analyses were performed using t-test; HaCaT, P = 0.006 at 24h and P = 0.002 at 48h; A431-AD, P<0.0001 at 24h and at 48h. Bottom panel: Representative Western blot analyses of PC-PLC protein expression performed in total lysates of cells cultured in the presence or absence of 50 μg/ml D609 (n = 3 independent experiments); t0 = untreated cells at 72 hours after seeding; β actin was used as quantitative loading control.</p

ET-18-OCH₃ prevents CCL2 transcript accumulation in the cytoplasm of MDM.

<p>(A) MDM were treated with ET-18-OCH₃ (10 µM) or left untreated (control). After 1 hour, some cultures were exposed to R5 gp120 (3 µg/ml) for 1 hour. Cells were then fixed, permeabilized, stained with rabbit anti-p65 Ab (showed in pseudocolor gray) and examined by CLSM. Nuclei are reported in blue (DAPI). Panels are representative of 4 independent experiments. The bars correspond to 20 µm. (B–C) MDM were treated with ET-18-OCH₃ (10 µM) for 1 hour prior to R5 gp120 (3 µg/ml) exposure for 4 hours. Total (B) or cytoplasmic (C) RNA was then extracted, retrotranscribed and amplified as described in Methods. The 2^∧(ΔΔCt) values for CCL2 transcripts were calculated as described in Methods. Data represent mean values (±SE) of the results obtained with MDM from 3 different donors. *p<0.05 (gp120 <i>vs</i> control; ET-18-OCH₃+gp120 <i>vs</i> gp120 alone).</p

CCR5 and G_iα inhibitors block R5 gp120-induced nuclear localization of PI-PLC β1 in MDM.

<p>(A) MDM were treated with PTX (10 ng/ml) for 2 hours or Tak779 (5 µM) for 1 hour prior to R5 gp120 (3 µg/ml) or CCL4 (200 ng/ml) exposure for 20 minutes. Cells were then fixed, permeabilized and stained with anti-PI-PLC β1 Ab. The cellular distribution of PI-PLC β1 (pseudocolor gray) was monitored by CLSM analysis. Panels are representative of 4 (Tak779) and 3 (PTX) independent experiments. The bars correspond to 20 µm. (B) MDM were treated with PTX (10 ng/ml) for 2 hours prior to R5 gp120 (3 µg/ml) exposure. After 24 hours of culture, supernatants were harvested and frozen before CCL2 determination. Data represent mean values (±SE) of the results obtained with the 9 donors tested. **p<0.01 (PTX+gp120 <i>vs</i> gp120 alone); ***p<0.001 (gp120 <i>vs</i> control).</p

ET-18-OCH₃ does not affect gp120-induced PC-PLC cellular relocalization in MDM.

<p>MDM were treated with ET-18-OCH₃ (10 µM) or left untreated (control). After 1 hour, some cultures were exposed to R5 gp120 (3 µg/ml) for 5 minutes. Cells were then fixed, permeabilized, stained with rabbit anti-PC-PLC Ab (pseudocolor gray) and examined by CLSM. Nuclei are reported in blue (DAPI). Representative examples of 3 independent experiments are shown. The bars correspond to 10 µm.</p

Effects of PC-PLC inhibition on EGFR, ERK and AKT phosphorylation in the HaCat and A431-AD cells.

<p>Representative Western blot analyses of total cell lysates from HaCaT (left panels) and A431-AD (right panels) cells cultured in the presence or absence of 50 μg/ml of D609. Cell lysates were immunoblotted with the following antibodies: pEGFR (Tyr1068), EGFR, pERK1/2 (Thr202/Tyr204), ERK 1/2, pAKT (Ser473), AKT and β-actin. β-actin was used as a quantitative loading control. Histograms below each panel represent the relative optical densities of phospho-protein levels normalized to the total protein level (mean values ± SD of three independent experiments) and are presented relative to the untreated sample at 24h. Statistical analyses were performed between treated samples (24h and 48 h) and untreated control sample at 24h, using the t-test.</p

HIV-1 gp120 induces nuclear localization of PI-PLC β1 in MDM.

<p>(A) MDM were treated with R5 gp120 (3 µg/ml) for the indicated time periods or left untreated (control). Cells were then fixed, permeabilized, stained with anti-PI-PLC β1 Ab (shown in pseudocolor gray) and examined by CLSM. Nuclei are reported in blue (DAPI). Panels are representative of 4 independent experiments. The bars correspond to 10 µm. (B) MDM were treated with R5 gp120 (3 µg/ml) for 20 minutes and then lysed. PI-PLC β1 expression in cytoplasmic and nuclear extracts was detected by western blotting analysis. Lamin A and β-actin were used as house-keeping control for nuclear and cytoplasmic fractions, respectively. The results from one representative experiment of 4 independently performed are shown. (C) Densitometric analyses of PI-PLC β1 expression performed on immunoblotting of MDM nuclear extracts from the 4 different donors analyzed. *p<0.05. (D) CLSM examinations of cells treated as described in A and then fixed, permeabilized and stained with anti-PI-PLC γ1 Ab (shown in pseudocolor gray). Panels are representative of 4 independent experiments. The bars correspond to 10 µm.</p

R5 gp120-activated ERK 1/2 mediates nuclear localization of PI-PLC β1 and CCL2 secretion in MDM.

<p>(A) MDM were treated with R5 gp120 (3 µg/ml) for the indicated time periods or left untreated (control). Cells were then lysed and ERK 1/2 phosphorylation was detected by western blotting. The results from one representative experiment of 4 independently performed are shown. (B) MDM were treated as described in A and ERK 1/2 phosphorylation was analyzed by flow cytometry after 10 minutes of gp120 exposure. Blue and red lines represent control and gp120-treated cells, respectively. Black line represents background staining of isotype-matched control Ab. The results from one representative experiment of 4 independently performed are shown. (C) MDM were treated with PD98059 (10 µM) for 1 hour prior to R5 gp120 (3 µg/ml) or CCL4 (200 ng/ml) exposure for 20 minutes. Cells were then fixed, permeabilized, stained with anti-PI-PLC β1 Ab (pseudocolor gray) and examined by CLSM. Representative examples of 5 independent experiments are shown. The bars correspond to 20 µm. (D) MDM were treated with PD98059, U0126, U0124 (10 µM) or left untreated (control). After 1 hour, some cultures were exposed to R5 gp120 (3 µg/ml). After 24 hours of culture, supernatants were harvested and frozen before CCL2 determination. Data represent mean values (±SE) of the results obtained with MDM from 10 different donors. *p<0.05; ***p<0.001.</p