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Mingzhao Xing - One of the best experts on this subject based on the ideXlab platform.
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Western blotting analysis of NIS protein expression in M14, HepG2 and MKN-7 cells after treatment with the MEK, Akt and HDAC inhibitors.
2013Co-Authors: Zhi Liu, Mingzhao XingAbstract:Cells were treated for 30 hrs with combination use of RDEA119, Perifosine, and SAHA at the concentrations described in Fig. 1, followed by standard Western blotting analysis of cell lysates using the specific primary antibody against NIS. The expression level of ß-actin was analyzed in parallel for quality control. “Control”, treatment of cells with vehicle; “R+P+S”, treatment of cells with combined use of RDEA119, Perifosine and SAHA.
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Target effects of inhibitors of the MAP kinase and PI3K/Akt pathways and HDAC in various non-thyroid human cancer cells.
2013Co-Authors: Zhi Liu, Mingzhao XingAbstract:The MEK inhibitor RDEA119, the Akt inhibitor Perifosine, and the HDAC inhibitor SAHA were used to respectively target these signaling pathways or molecules. Cells were treated for 30 hrs with 0.5 µM RDEA119, 5 µM Perifosine or 0.5 µM SAHA as indicated. DMSO or PBS was used in parallel as the vehicle control. Cells were lysed for Western blotting after treatments to reveal the levels of phosphorylated ERK (p-ERK), phosphorylated Akt (p-Akt), and acetylated histone (Ac-His) with specific antibodies. “+”, treatment with the indicated inhibitor; “−”, treatment with vehicle.
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Induction of radioiodine uptake in M14, HepG2 and MKN-7 cells by treatment with the MEK, Akt and HDAC inhibitors.
2013Co-Authors: Zhi Liu, Mingzhao XingAbstract:Cells were treated with RDEA119, Perifosine and SAHA as described in Fig. 2, followed by incubation with Na125I for 1 hr. Parallel cells were additionally treated with the NIS blocker NaClO4 to obtain non-specific radioiodine uptake/binding with the cells. Cells were then washed, lysed, and measured for radioactivity. Cell radioiodine uptake is presented as cpm/106 cells (on the y-axis of the figure) after correction for the non-specific radioiodine binding. Detailed experimental procedures are as described in the Materials and Methods. “Control”, treatment of cells with vehicle; “R+P+S”, treatment of cells with combination use of RDEA119, Perifosine and SAHA. ** In comparison with control, p
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NIS mRNA expression in different human cancer cells after drug treatments (mean±SD).*
2013Co-Authors: Zhi Liu, Mingzhao XingAbstract:*The data in the table represent folds of increase in NIS expression over the control treatment with DMSO/PBS as described in the Materials and Methods. RD, RDEA119; PE, Perifosine; SA, SAHA.
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induction of sodium iodide symporter nis expression and radioiodine uptake in non thyroid cancer cells
PLOS ONE, 2012Co-Authors: Zhi Liu, Mingzhao XingAbstract:Background This study was designed to explore the therapeutic potential of suppressing MAP kinase and PI3K/Akt pathways and histone deacetylase (HDAC) to induce the expression of sodium/iodide symporter (NIS) and radioiodine uptake in non-thyroid cancer cells. Methods We tested the effects of the MEK inhibitor RDEA119, the Akt inhibitor Perifosine, and the HDAC inhibitor SAHA on NIS expression in thirteen human cancer cell lines derived from melanoma, hepatic carcinoma, gastric carcinoma, colon carcinoma, breast carcinoma, and brain cancers. We also examined radioiodine uptake and histone acetylation at the NIS promoter in selected cells. Results Overall, the three inhibitors could induce NIS expression, to various extents, in melanoma and all the epithelial carcinoma-derived cells but not in brain cancer-derived cells. SAHA was most effective and its effect could be significantly enhanced by RDEA119 and Perifosine. The expression of NIS, at both mRNA and protein levels, was most robust in the melanoma cell M14, hepatic carcinoma cell HepG2, and the gastric carcinoma cell MKN-7 cell. Radioiodine uptake was correspondingly induced, accompanied by robust increase in histone acetylation at the NIS promoter, in these cells when treated with the three inhibitors. Conclusions This is the first demonstration that simultaneously suppressing the MAP kinase and PI3K/Akt pathways and HDAC could induce robust NIS expression and radioiodine uptake in certain non-thyroid human cancer cells, providing novel therapeutic implications for adjunct radioiodine treatment of these cancers.
Zhi Liu - One of the best experts on this subject based on the ideXlab platform.
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Western blotting analysis of NIS protein expression in M14, HepG2 and MKN-7 cells after treatment with the MEK, Akt and HDAC inhibitors.
2013Co-Authors: Zhi Liu, Mingzhao XingAbstract:Cells were treated for 30 hrs with combination use of RDEA119, Perifosine, and SAHA at the concentrations described in Fig. 1, followed by standard Western blotting analysis of cell lysates using the specific primary antibody against NIS. The expression level of ß-actin was analyzed in parallel for quality control. “Control”, treatment of cells with vehicle; “R+P+S”, treatment of cells with combined use of RDEA119, Perifosine and SAHA.
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Target effects of inhibitors of the MAP kinase and PI3K/Akt pathways and HDAC in various non-thyroid human cancer cells.
2013Co-Authors: Zhi Liu, Mingzhao XingAbstract:The MEK inhibitor RDEA119, the Akt inhibitor Perifosine, and the HDAC inhibitor SAHA were used to respectively target these signaling pathways or molecules. Cells were treated for 30 hrs with 0.5 µM RDEA119, 5 µM Perifosine or 0.5 µM SAHA as indicated. DMSO or PBS was used in parallel as the vehicle control. Cells were lysed for Western blotting after treatments to reveal the levels of phosphorylated ERK (p-ERK), phosphorylated Akt (p-Akt), and acetylated histone (Ac-His) with specific antibodies. “+”, treatment with the indicated inhibitor; “−”, treatment with vehicle.
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Induction of radioiodine uptake in M14, HepG2 and MKN-7 cells by treatment with the MEK, Akt and HDAC inhibitors.
2013Co-Authors: Zhi Liu, Mingzhao XingAbstract:Cells were treated with RDEA119, Perifosine and SAHA as described in Fig. 2, followed by incubation with Na125I for 1 hr. Parallel cells were additionally treated with the NIS blocker NaClO4 to obtain non-specific radioiodine uptake/binding with the cells. Cells were then washed, lysed, and measured for radioactivity. Cell radioiodine uptake is presented as cpm/106 cells (on the y-axis of the figure) after correction for the non-specific radioiodine binding. Detailed experimental procedures are as described in the Materials and Methods. “Control”, treatment of cells with vehicle; “R+P+S”, treatment of cells with combination use of RDEA119, Perifosine and SAHA. ** In comparison with control, p
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NIS mRNA expression in different human cancer cells after drug treatments (mean±SD).*
2013Co-Authors: Zhi Liu, Mingzhao XingAbstract:*The data in the table represent folds of increase in NIS expression over the control treatment with DMSO/PBS as described in the Materials and Methods. RD, RDEA119; PE, Perifosine; SA, SAHA.
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induction of sodium iodide symporter nis expression and radioiodine uptake in non thyroid cancer cells
PLOS ONE, 2012Co-Authors: Zhi Liu, Mingzhao XingAbstract:Background This study was designed to explore the therapeutic potential of suppressing MAP kinase and PI3K/Akt pathways and histone deacetylase (HDAC) to induce the expression of sodium/iodide symporter (NIS) and radioiodine uptake in non-thyroid cancer cells. Methods We tested the effects of the MEK inhibitor RDEA119, the Akt inhibitor Perifosine, and the HDAC inhibitor SAHA on NIS expression in thirteen human cancer cell lines derived from melanoma, hepatic carcinoma, gastric carcinoma, colon carcinoma, breast carcinoma, and brain cancers. We also examined radioiodine uptake and histone acetylation at the NIS promoter in selected cells. Results Overall, the three inhibitors could induce NIS expression, to various extents, in melanoma and all the epithelial carcinoma-derived cells but not in brain cancer-derived cells. SAHA was most effective and its effect could be significantly enhanced by RDEA119 and Perifosine. The expression of NIS, at both mRNA and protein levels, was most robust in the melanoma cell M14, hepatic carcinoma cell HepG2, and the gastric carcinoma cell MKN-7 cell. Radioiodine uptake was correspondingly induced, accompanied by robust increase in histone acetylation at the NIS promoter, in these cells when treated with the three inhibitors. Conclusions This is the first demonstration that simultaneously suppressing the MAP kinase and PI3K/Akt pathways and HDAC could induce robust NIS expression and radioiodine uptake in certain non-thyroid human cancer cells, providing novel therapeutic implications for adjunct radioiodine treatment of these cancers.
Mark M Souweidane - One of the best experts on this subject based on the ideXlab platform.
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dual inhibition of pi3k akt and mek erk pathways induces synergistic antitumor effects in diffuse intrinsic pontine glioma cells
Translational Oncology, 2017Co-Authors: Uday Bhanu Maachani, Melanie Schweitzer, Ranjodh Singh, Melinda Wang, Raymond Chang, Mark M SouweidaneAbstract:Diffuse intrinsic pontine glioma (DIPG) is a devastating disease with an extremely poor prognosis. Recent studies have shown that platelet-derived growth factor receptor (PDGFR) and its downstream effector pathway, PI3K/AKT/mTOR, are frequently amplified in DIPG, and potential therapies targeting this pathway have emerged. However, the addition of targeted single agents has not been found to improve clinical outcomes in DIPG, and targeting this pathway alone has produced insufficient clinical responses in multiple malignancies investigated, including lung, endometrial, and bladder cancers. Acquired resistance also seems inevitable. Activation of the Ras/Raf/MEK/ERK pathway, which shares many nodes of cross talk with the PI3K/AKT pathway, has been implicated in the development of resistance. In the present study, Perifosine, a PI3K/AKT pathway inhibitor, and trametinib, a MEK inhibitor, were combined, and their therapeutic efficacy on DIPG cells was assessed. Growth delay assays were performed with each drug individually or in combination. Here, we show that dual inhibition of PI3K/AKT and MEK/ERK pathways synergistically reduced cell viability. We also reveal that trametinib induced AKT phosphorylation in DIPG cells that could not be effectively attenuated by the addition of Perifosine, likely due to the activation of other compensatory mechanisms. The synergistic reduction in cell viability was through the pronounced induction of apoptosis, with some effect from cell cycle arrest. We conclude that the concurrent inhibition of the PI3K/AKT and MEK/ERK pathways may be a potential therapeutic strategy for DIPG.
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Dual Inhibition of PI3K/AKT and MEK/ERK Pathways Induces Synergistic Antitumor Effects in Diffuse Intrinsic Pontine Glioma Cells
Elsevier, 2017Co-Authors: Uday Bhanu Maachani, Melanie Schweitzer, Ranjodh Singh, Melinda Wang, Raymond Chang, Mark M SouweidaneAbstract:Diffuse intrinsic pontine glioma (DIPG) is a devastating disease with an extremely poor prognosis. Recent studies have shown that platelet-derived growth factor receptor (PDGFR) and its downstream effector pathway, PI3K/AKT/mTOR, are frequently amplified in DIPG, and potential therapies targeting this pathway have emerged. However, the addition of targeted single agents has not been found to improve clinical outcomes in DIPG, and targeting this pathway alone has produced insufficient clinical responses in multiple malignancies investigated, including lung, endometrial, and bladder cancers. Acquired resistance also seems inevitable. Activation of the Ras/Raf/MEK/ERK pathway, which shares many nodes of cross talk with the PI3K/AKT pathway, has been implicated in the development of resistance. In the present study, Perifosine, a PI3K/AKT pathway inhibitor, and trametinib, a MEK inhibitor, were combined, and their therapeutic efficacy on DIPG cells was assessed. Growth delay assays were performed with each drug individually or in combination. Here, we show that dual inhibition of PI3K/AKT and MEK/ERK pathways synergistically reduced cell viability. We also reveal that trametinib induced AKT phosphorylation in DIPG cells that could not be effectively attenuated by the addition of Perifosine, likely due to the activation of other compensatory mechanisms. The synergistic reduction in cell viability was through the pronounced induction of apoptosis, with some effect from cell cycle arrest. We conclude that the concurrent inhibition of the PI3K/AKT and MEK/ERK pathways may be a potential therapeutic strategy for DIPG
Mohamed Rahmani - One of the best experts on this subject based on the ideXlab platform.
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the bh3 only protein bim plays a critical role in leukemia cell death triggered by concomitant inhibition of the pi3k akt and mek erk1 2 pathways
Blood, 2009Co-Authors: Mohamed Rahmani, Anh Anderson, Joseph Habibi, Timothy Ryan Crabtree, Paul Dent, Mandy Mayo, Hisashi Harada, Andrea Ferreiragonzalez, Steven GrantAbstract:Mechanisms underlying apoptosis induced by concomitant interruption of the mitogen-activated protein kinase kinase/extracellular signal-regulated kinase 1/2 (MEK/ERK1/2) and phosphatidylinositol 3-kinase (PI3K)/Akt pathways were investigated in human leukemia cells. Inhibition of these pathways using the MEK inhibitor PD184352 or U0126 and the PI3K/Akt inhibitor Perifosine strikingly induced apoptosis in multiple malignant human hematopoietic cells, and substantially reduced the colony-forming capacity of primary acute myeloblastic leukemia, but not normal CD34+ cells. These events were associated with pronounced Bim up-regulation, Mcl-1 down-regulation, marked Bak/Bax conformational change accompanied by Bax membrane translocation, and a pronounced increase in Bax/Bak association. Molecular studies using tet-inducible Akt, constitutively active MEK1, dominant-negative Akt, and MEK1 small interfering RNA revealed that inhibition of both MEK/ERK1/2 and Akt pathways plays a critical functional role in Perifosine/PD184352-mediated lethality. Ectopic Mcl-1 expression potently inhibited Perifosine/PD184352-induced apoptosis, as did Bak or Bax knockdown. Notably, knockdown of Bim, but not Bad, blocked Bak and Bax conformational change, inhibited Bax membrane translocation, diminished Bax/Bak binding, and sharply attenuated Perifosine/PD184352-induced apoptosis. Finally, enforced expression of Bim significantly enhanced apoptosis induced by PI3K/Akt inhibitors, analogous to the effects of MEK1/2 inhibitors. Collectively, these findings suggest that Bim, and Mcl-1, but not Bad, integrate death signaling triggered by concomitant disruption of the PI3K/Akt and MEK1/2/ERK1/2 pathways in human leukemia cells.
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the bh3 only protein bim plays a critical role in cell death triggered by concomitant inhibition of the pi3k akt and mek erk pathways in human leukemia cells
Cancer Research, 2008Co-Authors: Mohamed Rahmani, Anh Anderson, Joseph Habibi, Timothy Ryan Crabtree, Paul Dent, Steven GrantAbstract:2332 Mechanisms underlying apoptosis induced by concomitant interruption of the MEK/ERK1/2 and Akt pathways were investigated in human leukemia cells. Apoptosis induction by Perifosine, an alkyl-lysophospholipid (ALP) that inhibits PI3K/Akt and activates ERK1/2, was dramatically potentiated by co-administration of the MEK1/2 inhibitors PD184352 or U0126 in multiple malignant hematopoietic cells. These, events were associated with pronounced JNK1/2 activation, Mcl-1 downregulation, Bim upregulation, and marked Bak/Bax conformational change accompanied by Bax membrane translocation. Enforced expression of constitutively active (myristoylated) Akt sharply reduced Perifosine/PD184352-mediated lethality. Genetic or pharmacological inhibition of JNK activation by shRNA directed against JNK1/2 or by the JNK inhibitor SP600125 significantly attenuated PD184352/Perifosine-induced apoptosis, but not Mcl-1 downregulation. Notably, ectopic Mcl-1 expression potently inhibited Perifosine/PD184352 lethality, as did Bak or Bax knockdown.Notably, knockdown of Bim, but not Bad, with shRNA blocked Bak and Bax conformational change, Bax membrane translocation, and apoptosis induced by Perifosine/PD184352. Conversely, enforced expression of mutant Bim (ser55,65,100→ala) that is unphosphorylatable by ERK1/2 significantly enhanced apoptosis induced by PI3/Akt kinase inhibitors (i.e. Perifosine, LY294002, PI-103). Collectively, these findings suggest that Bim, Mcl-1, and JNK, but not Bad, integrate death signaling triggered by concomitant disruption of the PI3K/Akt and MEK1/2/ERK1/2 pathways in human leukemia cells.
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coadministration of histone deacetylase inhibitors and Perifosine synergistically induces apoptosis in human leukemia cells through akt and erk1 2 inactivation and the generation of ceramide and reactive oxygen species
Cancer Research, 2005Co-Authors: Mohamed Rahmani, Eri Reese, Yu Dai, Cheryl Aue, Shaw G Payne, Sarah Spiegel, Steve GraAbstract:Interactions between histone deacetylase inhibitors (HDACIs) and the alkyl-lysophospholipid Perifosine were examined in human leukemia cells. Coadministration of sodium butyrate, suberoylanilide hydroxamic acid (SAHA), or trichostatin with Perifosine synergistically induced mitochondrial dysfunction (cytochrome c and apoptosis-inducing factor release), caspase-3 and -8 activation, apoptosis, and a marked decrease in cell growth in U937 as well as HL-60 and Jurkat leukemia cells. These events were associated with inactivation of extracellular signal-regulated kinase (ERK) 1/2 and Akt, p46 c-jun-NH2-kinase (JNK) activation, and a pronounced increase in generation of ceramide and reactive oxygen species (ROS). They were also associated with up-regulation of Bak and a marked conformational change in Bax accompanied by membrane translocation. Ectopic expression of Bcl-2 delayed but was ultimately ineffective in preventing Perifosine/HDACI-mediated apoptosis. Enforced expression of constitutively active mitogen-activated protein kinase kinase (MEK) 1 or myristoylated Akt blocked HDACI/Perifosine-mediated ceramide production and cell death, suggesting that MEK/ERK and Akt inactivation play a primary role in these phenomena. However, inhibition of JNK activation (e.g., by the JNK inhibitor SP600125) did not attenuate sodium butyrate/Perifosine-induced apoptosis. In addition, the free radical scavenger N-acetyl-L-cysteine attenuated ROS generation and apoptosis mediated by combined treatment. Finally, the acidic sphingomyelinase inhibitor desipramine attenuated HDACI/Perifosine-mediated ceramide and ROS production as well as cell death. Together, these findings indicate that coadministration of HDACIs with Perifosine in human leukemia cells leads to Akt and MEK/ERK disruption, a marked increase in ceramide and ROS production, and a striking increase in mitochondrial injury and apoptosis. They also raise the possibility that combining these agents may represent a novel antileukemic strategy.
Laura Moro - One of the best experts on this subject based on the ideXlab platform.
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Perifosine as a potential novel anti-cancer agent inhibits EGFR/MET-AKT axis in malignant pleural mesothelioma. PlosOne
2016Co-Authors: Giulia Pinton, Arcangela Gabriella Manente, Giovanni Angeli, Luciano Mutti, Laura MoroAbstract:Background: PI3K/AKT signalling pathway is aberrantly active and plays a critical role for cell cycle progression of human malignant pleural mesothelioma (MMe) cells. AKT is one of the important cellular targets of Perifosine, a novel bio-available alkylphospholipid that has displayed significant anti-proliferative activity in vitro and in vivo in several human tumour model systems and is currently being tested in clinical trials. Methods: We tested Perifosine activity on human mesothelial cells and different mesothelioma cell lines, in order to provide evidence of its efficacy as single agent and combined therapy. Results: We demonstrate here that Perifosine, currently being evaluated as an anti-cancer agent in phase 1 and 2 clinical trials, caused a dose-dependent reduction of AKT activation, at concentrations causing MMe cell growth arrest. In this study we firstly describe that MMe cells express aside from AKT1 also AKT3 and that either the myristoylated, constitutively active, forms of the two proteins, abrogated Perifosine-mediated cell growth inhibition. Moreover, we describe here a novel mechanism of Perifosine that interferes, upstream of AKT, affecting EGFR and MET phosphorylation. Finally, we demonstrate a significant increase in cell toxicity when MMe cells were treated with Perifosine in combination with cisplatin. Conclusions: This study provides a novel mechanism of action of Perifosine, directly inhibiting EGFR/MET-AKT1/3 axis
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Perifosine as a Potential Novel Anti-Cancer Agent Inhibits EGFR/MET-AKT Axis in Malignant Pleural Mesothelioma
2012Co-Authors: Giulia Pinton, Arcangela Gabriella Manente, Giovanni Angeli, Luciano Mutti, Laura MoroAbstract:BackgroundPI3K/AKT signalling pathway is aberrantly active and plays a critical role for cell cycle progression of human malignant pleural mesothelioma (MMe) cells. AKT is one of the important cellular targets of Perifosine, a novel bio-available alkylphospholipid that has displayed significant anti-proliferative activity in vitro and in vivo in several human tumour model systems and is currently being tested in clinical trials. MethodsWe tested Perifosine activity on human mesothelial cells and different mesothelioma cell lines, in order to provide evidence of its efficacy as single agent and combined therapy. ResultsWe demonstrate here that Perifosine, currently being evaluated as an anti-cancer agent in phase 1 and 2 clinical trials, caused a dose-dependent reduction of AKT activation, at concentrations causing MMe cell growth arrest. In this study we firstly describe that MMe cells express aside from AKT1 also AKT3 and that either the myristoylated, constitutively active, forms of the two proteins, abrogated Perifosine-mediated cell growth inhibition. Moreover, we describe here a novel mechanism of Perifosine that interferes, upstream of AKT, affecting EGFR and MET phosphorylation. Finally, we demonstrate a significant increase in cell toxicity when MMe cells were treated with Perifosine in combination with cisplatin. ConclusionsThis study provides a novel mechanism of action of Perifosine, directly inhibiting EGFR/MET-AKT1/3 axis, providing a rationale for a novel translational approach to the treatment of MMe.
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Expression of AKTs and rescue of Perifosine effects by transfection with constitutive active Myr-AKTs.
2012Co-Authors: Giulia Pinton, Arcangela Gabriella Manente, Giovanni Angeli, Luciano Mutti, Laura MoroAbstract:A, AKT1, 2, 3 and actin as control mRNA expression in A549 and REN cells was evaluated by RT-PCR as reported in in “Material and Methods” section. B, AKT1 and 3 protein expression was evaluated by Western blot analysis in REN cells with isoform specific antibodies, tubulin blot confirmed equal loading. Representative of three independent experiments. C, REN cells were transfected with 1 µg/plate of plasmid coding for the constitutive activated form of AKT, Myr-AKT1 or 3. 24 hours post transfection REN cells were treated with 23 µM Perifosine for 1 hour, then were lysed and equal amounts of proteins were subjected to SDS-PAGE followed by immuno-blotting with specific antibodies to phosphorylated AKT, HA to confirm transfection and tubulin for equal loading. D, The effect of Perifosine on cells transfected with Myr- AKT1 or 3 was also assessed on cell proliferation. REN cells were transfected with 1 µg/plate of plasmid coding Myr-AKT1 or 3 and 24 hours post transfection were treated with 23 µM Perifosine for 24 hours. At the end of treatment viable cell count was determined. Points, means ± SD of three individual measurements. Perifosine markedly alters AKT membrane translocation, so we next asked whether forced expression of an exogenous Myr-AKT1 or Myr-AKT3 could abrogate the effect of Perifosine on AKT activating phosphorylations and cell growth. Contradictory data are available in literature on the capability of a constitutively active AKT to overcome Perifosine action. While in prostate cancer cells inhibition of AKT phosphorylation was substantially relieved by introduction of a Myr-AKT1, which bypassed the requirement for PH domain-mediated membrane recruitment, in myeloma cells it has been described that Perifosine overcome constitutive highly active AKT1 [25], [26]. We transiently transfected REN cells either with an HA-tagged Myr-AKT1 or Myr-AKT3 and treated them for 24 hours with the IC50 dose of Perifosine. Western-blot analysis reported in Figure 3C shows that transfection of both Myr-AKT1 and Myr-AKT3 overcomes Perifosine inhibition of AKT phosphorylation in REN cells; only a slight reduction of phosphorylation, probably due to the block of the endogenous proteins, was observed. Cell growth analysis reported in Figure 3D demonstrates that both transfected Myr-AKTs rescued Perifosine induced block of cell proliferation.
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Isobologram analysis of drug interactions.
2012Co-Authors: Giulia Pinton, Arcangela Gabriella Manente, Giovanni Angeli, Luciano Mutti, Laura MoroAbstract:A, B, C, Isobologram plots of the interactions between Perifosine and cisplatin or gemcitabine or pemetrexed compounds on REN cells. Cells (5×104) were plated in complete medium and allowed to adhere to the surface overnight. The plating medium was removed and replaced with medium containing various concentrations of Perifosine and the IC50 doses of the other drugs. To determine the nature of the interaction between Perifosine and cis-platinum or gemcitabine or pemetrexed, we counted surviving cells following 24 hours drug incubation. The diagonal line represents the isoeffect line of additivity. Each point is the mean determined from experiments performed in triplicate.
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Effects of Perifosine on EGFR and MET phosphorylation and signalling.
2012Co-Authors: Giulia Pinton, Arcangela Gabriella Manente, Giovanni Angeli, Luciano Mutti, Laura MoroAbstract:A, B, Exponentially growing REN cells were pre-treated with 23 µM Perifosine for 1 hour and then incubated 10 minutes with EGF (5 ng/ml) or HGF (5 ng/ml). At the end of experiment, cell lysates were prepared or immunoprecipitated and analyzed by immunoblotting with indicated antibodies. Results are representative of three different experiments. B, D, The effect of Perifosine was also assessed on growth factor mediated cell proliferation. REN cells were treated with 23 µM Perifosine 24 hours in the absence or in the presence of 5 ng/ml EGF or HGF. At the end of treatment viable cell count was determined. Points, means ± SD of three individual measurements.