Induction of Growth Inhibition and Apoptosis by TZD18, a Novel Dual Ligand for PPAR alpha/gamma, in Human Ph-Positive ALL Cells In Vitro.

Abstract Peroxisome proliferator-activated receptors (PPARs) are ligand activated nuclear hormone receptors which play key roles in the differentiation and lipid metabolism of adipocytes. Recent data frequently indicated that PPAR ligands are also implicated in the growth inhibition, differentiation and apoptosis induction of several human cancers with diverse tissues. We previously showed that Pioglitazone (PGZ), a specific PPARgamma ligand and a member of the approved thiazolidinedione (TZD) class of anti-diabetic drugs, inhibited growth and induced apoptosis of human ALL cell lines including Ph-positive ALL cells (Zang et. al., Leukemia Research, 28:387, 2004). In this study, effects of a novel dual ligand specific for PPARalpha/gamma, TZD18 (MERCK, NJ, USA), on Ph-positive ALL cell lines, BV173, SD1 and Sup-B15 were examined. We noted that treatment of these cells with TZD18 resulted in growth inhibition in a dose-dependent manner which was associated with a G1 to S cell cycle arrest. This growth inhibition was much stronger than that of PGZ. However, this effect seemed not to be meditated through activation of PPARalpha or PPARgamma, since antagonists of PPARalpha or gamma could not reverse it. By studying the key regulators of cell cycle progression, we found that the expression of the cyclin dependent kinase inhibitor (CDKI) p27kip1, but not that of p21cip1, was enhanced whereas the expression of c-myc, cyclin D2, and cyclin dependent kinase 2 and 4 (CDK2 and CDK4) was decreased when these cells were treated with TZD18. Therefore, upregulation of p27kip1 and downregulation of cyclin Ds and CDKs may account for the G1 cell cycle arrest. Furthermore, a remarkable apoptosis induction was found in Ph-positive ALL cells treated with this dual ligand as measured by cell-death ELISA. No obvious alteration of bcl-2 levels but an upregulation of bax were observed in apoptotic cells. An activation of caspase-8 and caspase-9 by this ligand was also noticed. Of clinical importance, TZD18 enhanced the cytotoxic effect of Imatinib, a specific therapeutic agent for Ph-positive ALL. Overall, our findings strongly suggest that TZD18 may offer a new therapeutic agent for treatment of Ph-positive ALL in an adjuvant setting. (This study was supported by grants from Deutsche Jose Carreras Leukaemie-Stiftung and Deutsche Forschungsgemeinschaft to EE)

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PI3K a Selective Inhibitor Induces Growth Inhibition In Mantle Cell Lymphoma

Blood ◽

10.1182/blood.v116.21.1812.1812 ◽

2010 ◽

Vol 116 (21) ◽

pp. 1812-1812

Author(s):

Yixin Zhou ◽

Linhua Jin ◽

Stefania Pittaluga ◽

Mark Raffeld ◽

Takashi Miida ◽

...

Keyword(s):

Cell Cycle ◽

Growth Inhibition ◽

Cyclin D1 ◽

Cell Cycle Arrest ◽

Cell Lines ◽

Pi3k Inhibitor ◽

Dependent Manner ◽

Pi3k Inhibitor Ly294002 ◽

Pi3k Inhibitors ◽

Cycle Arrest

Abstract Abstract 1812 Deregulation of the phosphatidylinositol 3-kinase (PI3K)-mediated signaling plays an important role in the development of cell proliferation of mantle cell lymphoma (MCL). The PI3K pathway activation in MCL has been shown to result from constitutive B cell receptor (BCR) activation which is directly mediated by the Class IA PI3K p110 isoforms (a, β, and d). However, their relative contribution in MCL is not fully understood. In this study, the activity and molecular mechanisms of isoform-selective PI3K inhibitors which target different isoforms of the p110-kDa subunit has been investigated. We utilized the isoform-selective PI3K inhibitors; PI3-Ka inhibitor IV (p110a), TGX115 (p110b), IC87114 (p110d) and the non-specific PI3K inhibitor LY294002 (all inhibitors were purchased commercially). The p110a and p110d but not p110b isoform protein expression was detected in all tested MCL cell lines (Granta 519, JVM-2, Z138, Jeko-1, MINO). PI3-Ka inhibitor IV as well as non-specific PI3K inhibitor LY294002 induced cell growth inhibition with dose-dependent manner (IC50 at 48 hrs; PI3-Ka inhibitor IV: 17.5 μM for Granta 519, 14.3 μM for Jeko-1, 16.5 μM for Z138, LY294002: 14.8 μM for Granta 519, 19.4 μM for Jeko-1, 15.0 μM for Z138, MTT test). However, neither IC87114 nor TGX115 showed significant cell growth inhibition up to 40mM. Low dose of PI3-Ka inhibitor IV (5 μM) or LY294002 (5 μM) induced G0/G1 cell cycle arrest (increase of G0/G1 phase: PI3-Ka inhibitor IV 17.9 % for Granta 519, 28.2 % for Jeko-1, LY294002 19.3 % for Granta 519, 14.5 % for Jeko-1), and the higher dose (10 μM) increased apoptosis(specific apoptosis: PI3-Ka inhibitor IV 10.8 % for Granta 519, 15.3 % for Jeko-1, LY294002 13.6 % for Granta 519, 19.6 % for Jeko-1). No induction of cell cycle arrest/apoptosis by IC87114 or TGX115 treatment was observed. We then tried to assess the inhibition of PI3K/Akt signaling activation by p110a and p110d inhibitors. PI3-Ka inhibitor IV (10 μM) completely diminished phosphorylated (p-) Akt in all cell lines analyzed. Further investigation with 1–10 μM PI3-Ka inhibitor IV or IC87114 in Granta 519 and Jeko-1 cells declared that 1 μM PI3-Ka inhibitor IV almost diminished p-Akt and p-S6rp in both cells. The phosphorylation level of other PI3K/Akt signaling downstream substrates, GSK3-b and 4E-BP1, were down-regulated in dose dependent manner. Recently, GSK3-b kinase has been shown to negatively regulate cell cycle progression through Cyclin D1 repression in MCL. We observed that PI3-Ka inhibitor IV decreased Cyclin D1 expression and active pRb which are responsible for G0/G1 cell cycle arrest. The treatment with IC87114 (10 μM) performed moderate decrease of p-Akt, p-S6rp, and p-4E-BP, while no change in the levels of p-GSK3-b, Cyclin D1, or p-pRb was observed in both Granta 519 and Jeko-1 cells. We also tested whether the combination of PI3-Ka inhibitor IV or IC87114 with the proteasome inhibitor bortezomib induces synergistic cytotoxicity in MCL. No synergistic anti-proliferative effect was observed in any of the MCL cell lines analyzed. These findings demonstrate that p110a may be the responsible Class IA PI3K isoform for the development of MCL cell proliferation, and p110a isoform-selective PI3K inhibitor but not p110d or p110b inhibitors may provide a better therapeutic index relative to pan-PI3K inhibitors. Disclosures: No relevant conflicts of interest to declare.

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Apoptosis Induction, Cell Cycle Arrest and in Vitro Anticancer Activity of Gonothalamin in a Cancer Cell Lines

Asian Pacific Journal of Cancer Prevention ◽

10.7314/apjcp.2012.13.10.5131 ◽

2012 ◽

Vol 13 (10) ◽

pp. 5131-5136 ◽

Cited By ~ 28

Author(s):

Aied M. Alabsi ◽

Rola Ali ◽

Abdul Manaf Ali ◽

Sami Abdo Radman Al-Dubai ◽

Hazlan Harun ◽

...

Keyword(s):

Cell Cycle ◽

Anticancer Activity ◽

Cell Cycle Arrest ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

Apoptosis Induction ◽

Cycle Arrest

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Glioblastoma Multiforme Stem Cell Cycle Arrest by Alkylaminophenol through the Modulation of EGFR and CSC Signaling Pathways

Cells ◽

10.3390/cells9030681 ◽

2020 ◽

Vol 9 (3) ◽

pp. 681 ◽

Cited By ~ 3

Author(s):

Phuong Doan ◽

Aliyu Musa ◽

Akshaya Murugesan ◽

Vili Sipilä ◽

Nuno R. Candeias ◽

...

Keyword(s):

Cell Cycle ◽

Stem Cell ◽

Glioblastoma Multiforme ◽

Cell Cycle Arrest ◽

Signaling Pathways ◽

Cell Lines ◽

Cell Population ◽

Significant Role ◽

Dependent Manner ◽

Cycle Arrest

Cancer stem cells (CSCs), a small subpopulation of cells existing in the tumor microenvironment promoting cell proliferation and growth. Targeting the stemness of the CSC population would offer a vital therapeutic opportunity. 3,4-Dihydroquinolin-1(2H)-yl)(p-tolyl)methyl)phenol (THTMP), a small synthetic phenol compound, is proposed to play a significant role in controlling the CSC proliferation and survival. We assessed the potential therapeutic effects of THTMP on glioblastoma multiforme (GBM) and its underlying mechanism in various signaling pathways. To fully comprehend the effect of THTMP on the CSCs, CD133+ GBM stem cell (GSC) and CD133- GBM Non-stem cancer cells (NSCC) population from LN229 and SNB19 cell lines was used. Cell cycle arrest, apoptosis assay and transcriptome analysis were performed for individual cell population. THTMP strongly inhibited NSCC and in a subtle way for GSC in a time-dependent manner and inhibit the resistance variants better than that of temozolomide (TMZ). THTMP arrest the CSC cell population at both G1/S and G2/M phase and induce ROS-mediated apoptosis. Gene expression profiling characterize THTMP as an inhibitor of the p53 signaling pathway causing DNA damage and cell cycle arrest in CSC population. We show that the THTMP majorly affects the EGFR and CSC signaling pathways. Specifically, modulation of key genes involved in Wnt, Notch and Hedgehog, revealed the significant role of THTMP in disrupting the CSCs’ stemness and functions. Moreover, THTMP inhibited cell growth, proliferation and metastasis of multiple mesenchymal patient-tissue derived GBM-cell lines. THTMP arrests GBM stem cell cycle through the modulation of EGFR and CSC signaling pathways.

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Degalactotigonin, a Steroidal Glycoside from Solanum nigrum, Induces Apoptosis and Cell Cycle Arrest via Inhibiting the EGFR Signaling Pathways in Pancreatic Cancer Cells

BioMed Research International ◽

10.1155/2018/3120972 ◽

2018 ◽

Vol 2018 ◽

pp. 1-9 ◽

Cited By ~ 5

Author(s):

Hoang Le Tuan Anh ◽

Phuong Thao Tran ◽

Do Thi Thao ◽

Duong Thu Trang ◽

Nguyen Hai Dang ◽

...

Keyword(s):

Cell Cycle ◽

Pancreatic Cancer ◽

Cell Cycle Arrest ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

Solanum Nigrum ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Cycle Arrest

Degalactotigonin (1) and three other steroidal compounds solasodine (2), O-acetyl solasodine (3), and soladulcoside A (4) were isolated from the methanolic extract of Solanum nigrum, and their chemical structures were elucidated by spectroscopic analyses. The isolated compounds were evaluated for cytotoxic activity against human pancreatic cancer cell lines (PANC1 and MIA-PaCa2) and lung cancer cell lines (A549, NCI-H1975, and NCI-H1299). Only degalactotigonin (1) showed potent cytotoxicity against these cancer cell lines. Compound 1 induced apoptosis in PANC1 and A549 cells. Further study on its mechanism of action in PANC1 cells demonstrated that 1 significantly inhibited EGF-induced proliferation and migration in a concentration-dependent manner. Treatment of PANC1 cells with degalactotigonin induced cell cycle arrest at G0/G1 phase. Compound 1 induced downregulation of cyclin D1 and upregulation of p21 in a time- and concentration-dependent manner and inhibited EGF-induced phosphorylation of EGFR, as well as activation of EGFR downstream signaling molecules such as Akt and ERK.

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Biglycan induced growth inhibition in pancreatic cancer cell lines is caused by cell cycle arrest in G1

Gastroenterology ◽

10.1016/s0016-5085(00)84294-8 ◽

2000 ◽

Vol 118 (4) ◽

pp. A541

Author(s):

Gerrit Sommer ◽

Martina Weimer ◽

Uli Lacher ◽

Claudia Ruhland ◽

Christoph Wenger ◽

...

Keyword(s):

Cell Cycle ◽

Pancreatic Cancer ◽

Growth Inhibition ◽

Cell Cycle Arrest ◽

Cell Lines ◽

Cancer Cell ◽

Pancreatic Cancer Cell ◽

Cancer Cell Lines ◽

Cycle Arrest

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MDM4 is an essential disease driver targeted by 1q gain in Burkitt lymphoma

10.1101/289363 ◽

2018 ◽

Author(s):

Jennifer Hüllein ◽

Mikołaj Słabicki ◽

Maciej Rosolowski ◽

Alexander Jethwa ◽

Stefan Habringer ◽

...

Keyword(s):

Cell Cycle ◽

Tumor Suppressor ◽

Cell Cycle Arrest ◽

Cell Lines ◽

Burkitt Lymphoma ◽

Mutant Cell ◽

Negative Regulator ◽

Dependent Manner ◽

Cycle Arrest ◽

1Q Gain

AbstractOncogenic MYC activation promotes cellular proliferation in Burkitt lymphoma (BL), but also induces cell cycle arrest and apoptosis mediated by TP53, a tumor suppressor gene that is mutated in 40% of BL cases. To identify therapeutic targets in BL, we investigated molecular dependencies in BL cell lines using RNAi-based, loss-of-function screening. By integrating genotypic and RNAi data, we identified a number of genotype-specific dependencies including the dependence of TCF3/ID3 mutant cell lines on TCF3 and of MYD88 mutant cell lines on TLR signaling. TP53 wild-type (TP53wt) BL were dependent on MDM4, a negative regulator of TP53. In BL cell lines, MDM4 knockdown induced cell cycle arrest and decreased tumor growth in a xenograft model in a p53-dependent manner, while small molecule inhibition of the MDM4-p53 interaction restored p53 activity resulting in cell cycle arrest. Consistent with the pathogenic effect of MDM4 upregulation in BL, we found that TP53wt BL samples were enriched for gain of chromosome 1q which includes the MDM4 locus. 1q gain was also enriched across non-BL cancer cell lines (n=789) without TP53 mutation (23% in TP53wt and 12% in TP53mut, p<0.001). In a set of 216 cell lines representing 19 cancer entities from the Achilles project, MDM4 was the strongest genetic dependency in TP53wt cell lines (p<0.001).Our findings show that in TP53wt BL, MDM4-mediated inhibition of p53 is a mechanism to evade cell cycle arrest. The data highlight the critical role of p53 as a tumor suppressor in BL, and identifies MDM4 as a key functional target of 1q gain in a wide range of cancers, which is therapeutically targetable.

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PI3KCA and PIM Inhibitors in Peripheral T-Cell Lymphomas

Blood ◽

10.1182/blood.v116.21.4917.4917 ◽

2010 ◽

Vol 116 (21) ◽

pp. 4917-4917

Author(s):

Esperanza Martin-Sanchez ◽

Socorro M. Rodriguez-Pinilla ◽

Luis Lombardia ◽

Margarita Sanchez-Beato ◽

Beatriz Dominguez-Gonzalez ◽

...

Keyword(s):

Gene Expression ◽

Cell Cycle ◽

T Cell ◽

Cell Cycle Arrest ◽

Cell Lines ◽

Gene Expression Signature ◽

Dependent Manner ◽

Expression Signature ◽

Cycle Arrest ◽

T Cell Lymphomas

Abstract Abstract 4917 T-cell lymphomas (TCL) are a heterogeneous group of aggressive malignancies lacking specific and efficient therapy. Unfortunately, there are neither animal models nor representative cell lines for most TCL types, making functional and pharmacogenomics studies even more difficult. PI3K and PIM are kinases involved in cell proliferation, frequently altered in human cancer that seems to play a critical role in T-cell development and activation. Genomic studies have identified PIK3CD subunit to be significantly associated with in activation of CD40, NF-kB and TCR-pathways. The aim of this project is to determine the efficiency of PI3K inhibitors (PI3Ki) and PIM inhibitors (PIMi) in TCL, looking for biomarkers of their mechanism of action and to identify markers that could identify responders from non-responders. Twenty PTCL and seven reactive lymph nodes were studied using gene expression microarrays. We performed an in silico analysis using the Connectivity Map program to identify drugs that could potentially reverse PTCL gene expression signature. Among them, several PI3K/mTOR inhibitors were found. A panel of 6 TCL cell lines belonging to different TCL subgroups were treated with 3 PI3Ki (LY294002, ETP-45658, GDC-0941) and one PIMi (ETP-39010). Functional studies were also done to establish the role of each of the targeted genes. In vitro studies showed that PI3Ki induced G1 cell cycle arrest in all cell lines, and apoptosis in a portion of them, in a time/dose-dependent manner. We also observed a decrease in the levels of pAKT(S473), pGSK3B(S9) and p-p70S6K(T389) after treatment. In addition, both the analysis of the PTCL gene expression signature as well as western blot studies on TCL cell lines has shown overexpression of PIM family genes, A decrease in cell viability, and a strong induction of apoptosis in all cell lines was seen after PIM inhibition, without cell cycle arrest. Several diagnostic and pharmacodynamic biomarkers of PIMi have been identified at the mRNA and protein level in both cell lines In conclusion, our results indicate that PI3Ki and PIMi are effective therapeutic approaches for TCLs, identifying potential markers for patient's stratification and pharmacodynamic assessment. Disclosures: No relevant conflicts of interest to declare.

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Antiproliferative and Apoptosis Induction Potential of the Methanolic Leaf Extract ofHolarrhena floribunda(G. Don)

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2015/756482 ◽

2015 ◽

Vol 2015 ◽

pp. 1-11 ◽

Cited By ~ 19

Author(s):

J. A. Badmus ◽

O. E. Ekpo ◽

A. A. Hussein ◽

M. Meyer ◽

D. C. Hiss

Keyword(s):

Cell Cycle ◽

Cell Cycle Arrest ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

Molecular Probe ◽

Blue Dye ◽

Apoptosis Induction ◽

Cycle Arrest ◽

Wide Range

Natural plant products with potent growth inhibition and apoptosis induction properties are extensively being investigated for their cancer chemopreventive potential.Holarrhena floribunda(HF) is used in a wide range of traditional medicine practices. The present study investigated the antiproliferative and apoptosis induction potential of methanolic leaf extracts of HF against breast (MCF-7), colorectal (HT-29), and cervical (HeLa) cancer cells relative to normal KMST-6 fibroblasts. The MTT assay in conjunction with the trypan blue dye exclusion and clonogenic assays were used to determine the effects of the extracts on the cells. Caspase activities were assayed with Caspase-Glo 3/7 and Caspase-9 kits. Apoptosis induction was monitored by flow cytometry using the APOPercentage and Annexin V-FITC kits. Reactive oxygen species (ROS) was measured using the fluorogenic molecular probe 5-(and-6)-chloromethyl-2′,7′-dichlorofluorescein diacetate acetyl ester and cell cycle arrest was detected with propidium iodide. Dose-response analyses of the extract showed greater sensitivity in cancer cell lines than in fibroblast controls. Induction of apoptosis, ROS, and cell cycle arrest were time- and dose-dependent for the cancer cell lines studied. These findings provide a basis for further studies on the isolation, characterization, and mechanistic evaluation of the bioactive compounds responsible for the antiproliferative activity of the plant extract.

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Optimized anti–tumor effects of anthracyclines plus Vinca alkaloids using a novel, mechanism-based application schedule

Blood ◽

10.1182/blood-2010-02-269811 ◽

2011 ◽

Vol 118 (23) ◽

pp. 6123-6131 ◽

Cited By ~ 18

Author(s):

Harald Ehrhardt ◽

David Schrembs ◽

Christian Moritz ◽

Franziska Wachter ◽

Subrata Haldar ◽

...

Keyword(s):

Cell Cycle ◽

Cell Cycle Arrest ◽

Cell Lines ◽

Childhood Leukemia ◽

Underlying Mechanism ◽

Dependent Manner ◽

Vinca Alkaloids ◽

Cycle Arrest ◽

Induced Apoptosis

Abstract Application of anthracyclines and Vinca alkaloids on the same day represents a hallmark of polychemotherapy protocols for hematopoietic malignancies. Here we show, for the first time, that both drugs might act most efficiently if they are applied on different days. Proof-of-concept studies in 18 cell lines revealed that anthracyclines inhibited cell death by Vinca alkaloids in 83% of cell lines. Importantly, in a preclinical mouse model, doxorubicin reduced the anti–tumor effect of vincristine. Both drugs acted in a sequence-dependent manner and the strongest anti–tumor effect was obtained if both drugs were applied on different days. Most notably for clinical relevance, in 34% of 35 fresh primary childhood leukemia cells tested in vitro, doxorubicin reduced the anti–tumor effect of vincristine. As underlying mechanism, doxorubicin activated p53, p53 induced cell-cycle arrest, and cell-cycle arrest disabled inactivation of antiapoptotic Bcl-2 family members by vincristine; therefore, vincristine was unable to activate downstream apoptosis signaling. As molecular proof, antagonism was rescued by knockdown of p53, whereas knockdown of cyclin A inhibited vincristine-induced apoptosis. Our data suggest evaluating anthracyclines and Vinca alkaloids on different days in future trials. Selecting drug combinations based on mechanistic understanding represents a novel conceptional strategy for potent polychemotherapy protocols.

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