51 research outputs found
Additional file 1 of Identification of potential molecular targets for the treatment of cluster 1 human pheochromocytoma and paraganglioma via comprehensive proteomic characterization
Additional File 1. Table of all identified proteins (xls file
Additional file 3 of Identification of potential molecular targets for the treatment of cluster 1 human pheochromocytoma and paraganglioma via comprehensive proteomic characterization
Additional File 3. Topology prediction of selected differentially expressed IMPs with mapped peptides identified by the individual Pitchfork method
Additional file 2 of Identification of potential molecular targets for the treatment of cluster 1 human pheochromocytoma and paraganglioma via comprehensive proteomic characterization
Additional File 2. Volacano plots for the Pitchfork method
Histolopathology.
<p>Histologic features of liver PHEO lesions stained with H&E 24 hours after <i>i.p</i>. vehicle alone (left) or with <i>i.p.</i> LB1 at 1.5 mg/kg plus TMZ by gavage at 80 mg/kg (right). Exposure to a single <i>i.p.</i> injection of vehicle showed a homogeneous field of healthy appearing tumor cells, whereas combination treatment resulted in extensive necrosis of tumor cells.</p
Effect of LB1 and TMZ on tumor cell cycle and apoptosis.
<p>(A) Cell cycle analysis of MPC cells in exponential growth exposed for 48 hours to vehicle alone; LB1 alone at 5 µM; TMZ at 50 µM and; LB1 at 5 µM and TMZ at 50 µM. (B) PARP expression changes in 24 hours after treatment of mice bearing hepatic tumors with vehicle, LB1 alone at 1.5 mg/kg by gavage, TMZ alone by gavage at 80 mg/kg, of both drugs at the same doses.</p
Western blot on PHEO tumors.
<p>Changes in the state of phosphorylation and abundance of small pAKT, p53, pMDM2, and pPlk-1 24 hours after treatment of mice bearing hepatic tumors with vehicle, LB1 alone at 1.5 mg/kg by gavage, TMZ alone by gavage at 80 mg/kg, of both drugs at the same doses. (A) Western blots show that LB1 exposure increases pAKT in control-untreated tumors and treated tumors. TMZ does not change the expression of pAKT and combination of LB1 plus TMZ highly increases pAKT expression. (B) Western blots demonstrate marked increased expression of p53 after TMZ but complete inhibition of this induction by exposure to LB1 accompanied by an increase in the expression of pMDM2 in tumor cells exposed to LB1 alone or in combination with TMZ (C) Expression of pPlk1 shows a marked increase in tumors after exposure to the combination of drugs.</p
Western blot on MPC cells.
<p>Changes in pAKT, p53, pMDM2, and pPlk-1 after 24 hours treatment on MPC cells with 5 µM of LB1, 50 µM of TMZ and combination of both drugs. (A) Western blots show that LB1 exposure increases pAKT compared to control MPC cells (untreated, only vehicle). TMZ does not noticeably change the expression of pAKT and combination of LB1 and TMZ highly increases pAKT expression. (B) A demonstration of markedly increased expression of p53 after TMZ treatment but inhibition of expression by exposure to LB1. Noticeable increase in the expression of pMDM2 in MPC cells treated with LB1 alone or in combination with TMZ. (C) Noticeable increases in expression of pPlk1 in MPC cells after exposure to the combination of drugs.</p
<i>In vivo</i> anti-tumor activity of LB1 and TMZ and histological examination.
<p>Effects of treatment upon growth and molecular changes in hepatic tumors: (A) MRI images of untreated mice at weeks 5, 6, and 7 following intravenous injection of MPC cells and a photomicrograph showing the growth hepatic lesions and Alzet minipump. Barbed arrow indicates the gall bladder. Plain arrows indicate the same tumor nodules over time. (B) Inhibition of total hepatic tumor volume by LB1 alone at 1.5 mg/kg daily for 14 days by <i>c.i.</i> administered at 5<sup>th</sup> day after MPC cells injection; TMZ alone at 80 mg/kg for 3 doses administered at 15<sup>th</sup> day after MPC cells injection; and, combination of both drugs. (C) Survival curve combining the data from the study depicted in (B) a total of 10 control animals, 10 animals with combination treatment LB1 and TMZ, 5 animals with TMZ alone, and 5 animals with LB1 alone. Kaplan-Meier analysis revealed that survival following LB1 plus TMZ was significantly greater than with LB1 alone and TMZ alone (log rank, <i>P</i><0.0001). (D) MRI images of mice, treated with the combination of LB1 by <i>c.i</i>. for 14 days and 3 doses of TMZ, at week 7, 9, and 12. Partial response of treatment is presented with delayed appearance of hepatic tumors compared to untreated group. Complete response presents absence of hepatic tumors after treatment. A photomicrograph of the liver of one treated animal at week 12, showing the absence of gross tumor, and the presence of fibrous scar tissue (arrows). (E) Inhibition of estimated total hepatic tumor volume by combination treatment of LB1 and TMZ. LB1 at 1.5 mg/kg daily for 14 days by <i>c.i</i>. administered at 5<sup>th</sup> day after MPC cells injection and TMZ at 80 mg/kg every 3 days for 14 doses beginning on 15<sup>th</sup> day after MPC cells injection (with combination or alone). (F) Survival curve combining the data described in E. Total of 12 control animals, 5 animals for TMZ and 7 animals for combination treatment LB1 and TMZ. Survival of animals with combined treatment were significantly greater compared to controls (log rank, <i>P</i><0.0001). (G) Serial MRI images of mice, treated with the combination of LB1 and 14 doses TMZ with partial and complete responses. (H) Histologic features of liver PHEO at week 12 stained with H&E receiving no treatment or LB1 by <i>c.i.</i> and three doses of TMZ as described in D. Untreated animals showed intrahepatic deposits of cancer cells whereas the liver of an animal receiving both drugs that had no gross evidence of tumor revealed normal parenchyma and fibrous tissue, believed to be scarring at former sites of tumor masses. (I) Survival curves of animals treated with LB1 and 14 doses of TMZ when administration started at the same time, on day 5 after MPC cells injection. (n = 5 treated animals with LB1 plus TMZ, n = 5 for controls; log rank, <i>P</i> = 0.0035).</p
<i>In vitro</i> anti- proliferative activity of LB1 and TMZ and their combination.
<p>Inhibition of growth of MPC cells in culture: (A and B) Exposure for 3 days to increasing concentrations of LB1 or TMZ. (C–E) Exposure to increasing concentrations of LB1 plus TMZ. (F) Synergy analysis was done based on data from (A–E) using CalsuSyn software. CI values: C = 1 as additivity; C<1 as synergy; C>1 as antagonism. Combo 1 presents combination of 5 µM of LB1 and 100, 200, 300 µM of TMZ; Combo 2 presents combination of 7.5 µM of LB1 and 100, 200, 300 µM of TMZ; and Combo 3 10 µM of LB1 and 100, 200, 300 µM of TMZ.</p
Ganetespib inhibits tumor metastasis in a spontaneous metastatic pheochromocytoma model.
<p>A) Efficacy of ganetespib in an MTT-luc subcutaneous model of pheochromocytoma. Quantification of tumor growth was determined by luciferase measurement in an IVIS instrument. Data are plotted as mean photon counts (total flux) from regions of interest (ROI). Error bars represent standard deviations. B) Primary tumors excised from the two groups were compared. C and D) <i>Ex vivo</i> bioluminescence signals from lung and liver were compared between the two groups and graphed as total flux as a method of quantification of metastatic burden at the end of the experiment (*p<0.05).</p
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