MOLECULAR DOCKING OF SELECTED COMPOUNDS FROM CLINACANTHUS NUTANS WITH BCL-2, P53, CASPASE-3 AND CASPASE-8 PROTEINS IN THE APOPTOSIS PATHWAY

Abstract Background Influenza A virus (IAV) is still a major health threat. The clinical manifestations of this infection are related to immune dysregulation, which causes morbidity and mortality. The usage of traditional medication with immunomodulatory properties against influenza infection has been increased recently. Our previous study showed antiviral activity of quercetin-3-O-α-L-rhamnopyranoside (Q3R) isolated from Rapanea melanophloeos (RM) (L.) Mez (family Myrsinaceae) against H1N1 (A/PR/8/34) infection. This study aimed to confirm the wider range of immunomodulatory effect of Q3R on selective pro- and anti-inflammatory cytokines against IAV in vitro, to evaluate the effect of Q3R on apoptosis pathway in combination with H1N1, also to assess the physical interaction of Q3R with virus glycoproteins and RhoA protein using computational docking. Methods MDCK cells were exposed to Q3R and 100CCID50/100 μl of H1N1 in combined treatments (co-, pre- and post-penetration treatments). The treatments were tested for the cytokines evaluation at RNA and protein levels by qPCR and ELISA, respectively. In another set of treatment, apoptosis was examined by detecting RhoA GTPase protein and caspase-3 activity. Molecular docking was used as a tool for evaluation of the potential anti-influenza activity of Q3R. Results The expressions of cytokines in both genome and protein levels were significantly affected by Q3R treatment. It was shown that Q3R was much more effective against influenza when it was applied in co-penetration treatment. Q3R in combination with H1N1 increased caspase-3 activity while decreasing RhoA activation. The molecular docking results showed strong binding ability of Q3R with M2 transmembrane, Neuraminidase of 2009 pandemic H1N1, N1 and H1 of PR/8/1934 and Human RhoA proteins, with docking energy of − 10.81, − 10.47, − 9.52, − 9.24 and − 8.78 Kcal/mol, respectively. Conclusions Quercetin-3-O-α-L-rhamnopyranoside from RM was significantly effective against influenza infection by immunomodulatory properties, affecting the apoptosis pathway and binding ability to viral receptors M2 transmembrane and Neuraminidase of 2009 pandemic H1N1 and human RhoA cellular protein. Further research will focus on detecting the detailed specific mechanism of Q3R in virus-host interactions.

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Caspase-8 Regulates Caspase-3 and Rb Respectively in Fas- and Actinomycin D-Mediated Apoptosis Pathway in Human Hepatoma Bel-7402 Cells

2008 International Conference on BioMedical Engineering and Informatics ◽

10.1109/bmei.2008.146 ◽

2008 ◽

Author(s):

Yu Wang ◽

Liguang Sun ◽

Chunhui Xia ◽

Liping Ye ◽

Biao Wang

Keyword(s):

Caspase 3 ◽

Actinomycin D ◽

Caspase 8 ◽

Human Hepatoma ◽

Apoptosis Pathway

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Tetrandrine inhibits colon carcinoma HT-29 cells growth via the Bcl-2/Caspase 3/PARP pathway and G1/S phase

Bioscience Reports ◽

10.1042/bsr20182109 ◽

2019 ◽

Vol 39 (5) ◽

Cited By ~ 10

Author(s):

JiaNan Li ◽

QiuHong Wang ◽

ZhiBin Wang ◽

Na Cui ◽

BingYou Yang ◽

...

Keyword(s):

Colon Cancer ◽

Molecular Docking ◽

Cancer Cells ◽

Caspase 3 ◽

Caspase 8 ◽

Therapeutic Effects ◽

Dependent Manner ◽

Colon Cancer Cells ◽

Ht 29 ◽

Active Caspase

Abstract Tetrandrine (Tet) bisbenzylisoquinoline alkaloids isolated from Stephania tetrandra and other related species of Menispermaceae. It has been demonstrated to have positive therapeutic effects on cardiovascular disease, hypertension, silicosis, autoimmune diseases. In recent years, some reports have shown that Tet has anticancer activity in human cancers. To explore the pharmacological activity and mechanism of Tet on colon cancer and its unique advantages as a natural product. In the present study, analyses of the cell cycle, apoptosis, targets prediction, molecular docking, and alterations in protein levels were performed to elucidate how Tet functions in colon cancer. We found that Tet robustly induced arrest at the G1 phase in colon cancer cell line HT-29. It induced HT-29 cell apoptosis in a dose-dependent manner. Similarly, analysis of protein expression levels in HT-29 cells showed down-regulation of Bcl-2, pro-caspase 3, pro-caspase 8, PARP, cyclin D1 (CCND1), cyclin-dependent kinase 4 (CDK 4), and up-regulation of Bax, active caspase 3, and active caspase 8. These results indicate that Tet induces apoptosis of colon cancer cells through the mitochondrial pathway and caspase family pathway. Molecular docking showed interaction effects and binding energy. Comparing with the CDK4 inhibitors ribociclib and palbociclib, the docking energy is similar to the docked amino acid residues. Therefore, we conclude that Tet and the CCND1/CDK4 compound could form hydrogen bonds and a stable compound structure, which can inhibit colon cancer cells proliferation by regulating CCND1/CDK4 compound and its downstream proteins phosphorylated Rb (p-Rb). In summary, Tet may be a potential drug for colon cancer therapy.

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Smac Mimetic-Mediated Sensitization Of Childhood ALL Cells For Drug-Induced Apoptosis Independent Of TNFα and NFκB Signaling Pathways

Blood ◽

10.1182/blood.v122.21.2911.2911 ◽

2013 ◽

Vol 122 (21) ◽

pp. 2911-2911

Author(s):

Karin Schmelz ◽

Nina Weichert ◽

Jutta Proba ◽

Marie-Sophie Erdmann ◽

Patrick Hundsdoerfer

Keyword(s):

Signaling Pathways ◽

Tumor Cells ◽

Caspase 3 ◽

Caspase 8 ◽

Apoptosis Induction ◽

Caspase 9 ◽

Drug Induced ◽

Childhood All ◽

Apoptosis Pathway ◽

Induced Apoptosis

Abstract Targeting inhibitor of apoptosis proteins (IAPs) using small molecular Smac mimetics (SM) has been shown to offer a novel promising treatment strategy for resistant malignant diseases including childhood acute lymphoblastic leukemia (ALL). The effect of SM alone has been shown to be associated with endogenous TNFα expression, therefore tumor cells can be classified into sensitive or resistant against apoptosis induction by SM alone. In SM sensitive tumor cells the effect of SM has been shown to be mediated mainly by degradation of cellular IAP (cIAP) and activation of TNFα and NFκB signaling pathways but not inhibition of XIAP. We show here, that sensitivity of ALL cells to SM alone (as well as TNFα expression) is highly variable. Nevertheless even in ALL cells resistant against SM alone, treatment with SM resulted in significant sensitization for drugs used within standard induction therapy for childhood ALL. Sensitization for drug-induced apoptosis by SM was not only mediated by activation of the intrinsic (cleavage of caspase 9) but also extrinsic apoptosis pathway (cleavage of caspase 8). Surprisingly, SM-induced cIAP degradation alone was not sufficient for caspase 8 activation and apoptosis induction. Consistently, SM-mediated sensitization for drug-induced apoptosis was independent of TNFα and NFκB signaling pathways. We demonstrate that caspase 8 activation by combined treatment with SM and cytostatic drugs is blocked by inhibition of caspase 3 and caspase 9 and therefore occurs downstream of intrinsic apoptosis pathway activation. In conclusion, our data argue for a model comprising inhibition of XIAP-mediated blockade of caspase 3/9 as the central effect of SM in chemo-sensitization of childhood ALL cells resistant against SM-alone. Disclosures: No relevant conflicts of interest to declare.

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In vitro cytotoxicity of Clinacanthus nutans fractions on breast cancer cells and molecular docking study of sulphur containing compounds against caspase-3

Food and Chemical Toxicology ◽

10.1016/j.fct.2019.110869 ◽

2020 ◽

Vol 135 ◽

pp. 110869 ◽

Cited By ~ 2

Author(s):

Roziasyahira Mutazah ◽

Hazrulrizawati Abd Hamid ◽

Aizi Nor Mazila Ramli ◽

Mohd Fadhlizil Fasihi Mohd Aluwi ◽

Mashitah M. Yusoff

Keyword(s):

Breast Cancer ◽

Molecular Docking ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Caspase 3 ◽

Docking Study ◽

In Vitro Cytotoxicity ◽

Molecular Docking Study ◽

Clinacanthus Nutans

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A novel TNFR1-triggered apoptosis pathway mediated by class IA PI3Ks in neutrophils

Blood ◽

10.1182/blood-2010-11-322206 ◽

2011 ◽

Vol 117 (22) ◽

pp. 5953-5962 ◽

Cited By ~ 55

Author(s):

Barbara Geering ◽

Ursina Gurzeler ◽

Elena Federzoni ◽

Thomas Kaufmann ◽

Hans-Uwe Simon

Keyword(s):

Molecular Mechanisms ◽

Caspase 3 ◽

Caspase 8 ◽

Tnf Receptor ◽

Caspase Activation ◽

Apoptosis Pathway ◽

Tnf Α ◽

Pi3k Activation ◽

Amplification Loop ◽

Induced Apoptosis

Abstract The most common form of neutrophil death is apoptosis. In the present study, we report surprising differences in the molecular mechanisms used for caspase activation between FAS/CD95-stimulated and TNF receptor 1 (TNFR1)–stimulated neutrophils. Whereas FAS-induced apoptosis was followed by caspase-8 activation and required Bid to initiate the mitochondrial amplification loop, TNF-α–induced apoptosis involved class IA PI3Ks, which were activated by MAPK p38. TNF-α–induced PI3K activation resulted in the generation of reactive oxygen species, which activated caspase-3, a mechanism that did not operate in neutrophils without active NADPH oxidase. We conclude that in neutrophils, proapoptotic pathways after TNFR1 stimulation are initiated by p38 and PI3K, but not by caspase-8, a finding that should be considered in anti-inflammatory drug-development strategies.

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Caspase-8 Regulates Endoplasmic Reticulum Stress-Induced Necroptosis Independent of the Apoptosis Pathway in Auditory Cells

International Journal of Molecular Sciences ◽

10.3390/ijms20235896 ◽

2019 ◽

Vol 20 (23) ◽

pp. 5896 ◽

Cited By ~ 6

Author(s):

Kishino ◽

Hayashi ◽

Maeda ◽

Jike ◽

Hidai ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Cell Death ◽

Er Stress ◽

Caspase 3 ◽

Caspase 8 ◽

Intrinsic Apoptosis ◽

Caspase 9 ◽

Apoptosis Pathway ◽

Transmission Electron ◽

Initiator Caspases

The aim of this study is to elucidate the detailed mechanism of endoplasmic reticulum (ER) stress-induced auditory cell death based on the function of the initiator caspases and molecular complex of necroptosis. Here, we demonstrated that ER stress initiates not only caspase-9-dependent intrinsic apoptosis along with caspase-3, but also receptor-interacting serine/threonine kinase (RIPK)1-dependent necroptosis in auditory cells. We observed the ultrastructural characteristics of both apoptosis and necroptosis in tunicamycin-treated cells under transmission electron microscopy (TEM). We demonstrated that ER stress-induced necroptosis was dependent on the induction of RIPK1, negatively regulated by caspase-8 in auditory cells. Our data suggested that ER stress-induced intrinsic apoptosis depends on the induction of caspase-9 along with caspase-3 in auditory cells. The results of this study reveal that necroptosis could exist for the alternative backup cell death route of apoptosis in auditory cells under ER stress. Interestingly, our data results in a surge in the recognition that therapies aimed at the inner ear protection effect by caspase inhibitors like zVAD-fmk might arrest apoptosis but can also have the unanticipated effect of promoting necroptosis. Thus, RIPK1-dependent necroptosis would be a new therapeutic target for the treatment of sensorineural hearing loss due to ER stress.

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Investigation of Neuroprotective Potential of Anti-Diabetic Agents in Ischemic Brain Proteomics Through In-Silico Molecular Simulation Studies

Biointerface Research in Applied Chemistry ◽

10.33263/briac124.53475362 ◽

2021 ◽

Vol 12 (4) ◽

pp. 5347-5362

Keyword(s):

Molecular Docking ◽

Molecular Simulation ◽

In Silico ◽

Caspase 3 ◽

Caspase 8 ◽

Caspase 9 ◽

Discovery Studio ◽

Tnf Α ◽

Screening And Identification ◽

Neuroprotective Actions

Defective neurotransmission, impaired synaptic plasticity, and progressive neurodegeneration triggered by diabetes and stroke. The computational designing of a therapeutic agent, molecular dynamic (MD) simulation, and molecular docking are essential, along with streamlined and inexpensive techniques which enable the potential therapeutic targets for the novel drug discovery process in specific pathology. Molecular docking software such as AutoDock tool v1.5.6, PyRx v8.0, Discovery Studio Visualizer v19.1.0.18287, and PyMOL v2.3.1 has been used for virtual screening and identification of structural based molecular interactions. The 3D co-crystal structures of receptor/target therapeutic proteins (BCl-2, Caspase-3, Caspase-8, Caspase-9, IL6, MAPKERK, PI3K, TGF- β, TNF- α, and ZO-1) with attached ligand were gained from the RCSB-PDB website in PDB format. The obtained structures comprise a ligand and a water molecule, which must be removed before docking from the receptor using Discovery Studio Visualizer and finally transformed into macromolecules by AutoDock Vina of PyRx. The active residues were recognized through previously published literature. In-Silico molecular simulation study of anti-diabetic agents with stroke and neurodegeneration-associated proteins gave a strong hypothesis to the neuroprotective actions of the anti-diabetic drugs. Moreover, higher binding energy with all biomarker proteins directs us of exerting strong therapeutic action of the selected class of anti-diabetic agents, i.e., voglibose, saxagliptin, repaglinide, and Dapagliflozin with BCl-2, Caspase-3, Caspase-8, Caspase-9, IL6, MAPK/ERK, PI3K, TGF- β, TNF- α and ZO-1. The current study elucidates the strong anti-stroke potential of selected anti-diabetic agents and can claim a strong candidature as anti-stroke therapeutics in specific hyperglycemic conditions.

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109. TROPHOBLAST DEPORTATION IS DEPENDENT UPON CASPASES 3, 8 AND ROCK

Reproduction Fertility and Development ◽

10.1071/srb09abs109 ◽

2009 ◽

Vol 21 (9) ◽

pp. 28

Author(s):

K. J. Askelund ◽

P. Stone ◽

L. W. Chamley

Keyword(s):

Caspase 3 ◽

First Trimester ◽

Normal Pregnancy ◽

Maternal Blood ◽

Caspase 8 ◽

Caspase 9 ◽

Apoptotic Pathway ◽

Apoptosis Pathway ◽

Villous Trophoblast

Background: Trophoblast deportation is the process whereby multinucleated fragments of the syncytiotrophoblast are shed from the placenta into the maternal blood. It is estimated that 150,000 are shed from the placenta and deported daily in normal pregnancy and that more are shed during preeclampsia1. In normal pregnancy deported trophoblasts are thought to die by apoptosis, which is also increased in villous trophoblast in preeclampsia2. However, experimental confirmation that apoptosis leads to trophoblast shedding is required and it is not clear which components of the apoptotic pathway are involved in trophoblast shedding. Objectives: To determine the effect of inhibiting caspase 3 (executioner), caspases 8 and 9 (initiators), and Rho-associated kinase (ROCK; bleb formation) on the number of trophoblasts shed from first trimester human placentae. Methods : Using an in vitro placental explant model of trophoblast deportation, first trimester placentae were cultured for 72 hours in media containing specific inhibitors of ROCK, caspases 3, 8 or 9. Trophoblasts shed from quintuple explants/inhibitor from five placentae were depleted of contaminating leucocytes and erythrocytes, labelled with trypan blue and the sizes and numbers of shed trophoblasts quantified using a Nexcelom automated counter. Results: The number of trophoblasts that were shed from the explants was significantly increased (p=0.04) when caspase 3 (2.4 fold) and caspase 8 (2.7 fold) were inhibited. There was no significant change following caspase 9 inhibition. The number of shed trophoblasts was significantly decreased when ROCK was inhibited. None of the inhibitors significantly altered the size of the shed trophoblasts. Conclusion: Our data suggest that the apoptosis pathway is involved in trophoblast shedding in vitro from first trimester placentae. That caspase 8 but not caspase 9 affected shedding suggests trophoblasts from normal placentae are induced to die via the extrinsic apoptosis pathway. Aberrant regulation of the apoptosis pathway may contribute to pregnancy pathology.

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FLIPLong(L) and FLIPShort(S) Overexpression in the Human Myeloid Leukemia Cell Line ML-1 and Its Role in TRAIL [Tumor Necrosis Factor(TNF)-Related Apoptosis-Inducing Ligand] and TNFa Induced Apoptosis.

Blood ◽

10.1182/blood.v106.11.4378.4378 ◽

2005 ◽

Vol 106 (11) ◽

pp. 4378-4378

Author(s):

Sudeshna Seal ◽

Daniella B. Kerbauy ◽

Vladimir Lesnikov ◽

Nissa Abbasi-Shafer ◽

H. Joachim Deeg

Keyword(s):

Caspase 3 ◽

Leukemia Cell ◽

Leukemia Cell Line ◽

Caspase 8 ◽

Slight Reduction ◽

Western Blots ◽

Apoptosis Pathway ◽

Induced Apoptosis ◽

Active Caspase

Abstract TRAIL initiates activation of Caspase-8, which is blocked by the FLICE inhibitory protein (FLIP), resulting in resistance to apoptosis. Here we show that overexpression of FLIPL and FLIPS in ML1 cells, with low constitutive levels of FLIP, protects these cells against apoptosis induced by TRAIL, not only via Caspase-8 inhibition, but also via upregulation of anti-apoptotic molecules. Methods: 1) Apoptosis was determined by Annexin V/ 7-AAD following treatment with TRAIL (100–500ng/ml), or TNFa (20–100ng/ml) in ML1 cells transduced with FLIPL.GFP (green fluorescent protein), FLIPS.GFP or Neo.GFP (control). 2) Caspase-8, Caspase-3, Bid, Bcl-xL, XIAP, phosphorylated (P)-IKBa and P-Akt were determined by western blots. 3) Active Caspase-3 was determined using EnzChek Caspase-3 assay kit. Results: Both FLIPL and FLIPS transduction protected ML1 cells against apoptosis induced by TRAIL (300ng/ml), while no protection was observed in Neo.GFP cells. FLIPL had a more profound protective effect than FLIPS (Fig.1A). Both FLIPL and FLIPS, but not Neo.GFP, blocked Caspase-8 and Caspase-3 activation (Fig.1B); FLIPS cells showed two-fold higher levels of active Caspase-3 than FLIPL cells, consistent with higher apoptosis in FLIPS cells. Caspase-3 can be activated through Caspase-8 (extrinsic pathway) or via Caspase-8/Bid (intrinsic pathway). The latter was responsible for high active Caspase-3 levels in FLIPS cells as shown by the presence of cleaved Bid (t-Bid) (Fig.1B); cleavage of Bid was inhibited by combination of TRAIL and Z-IETD-FMK (Caspase-8 inhibitor). Anti-apoptotic molecules, including Bcl-xL, XIAP and FLIP are regulated by NF-kB and FLIP participates in an NF-kB auto-amplification loop. While Neo.GFP cells showed little Bcl-xL after 4h of TRAIL exposure and there was a twofold reduction in FLIPS cells, only a slight reduction of Bcl-xL was noted in FLIPL cells. FLIPL cells showed the lowest rates of apoptosis when exposed to TNFa and BMS543541, a specific inhibitor of IkB kinase (Fig. 1C). In the presence of BMS543541, phosphorylation of IkBa and levels of Bcl-xL and XIAP decreased in Neo.GFP and FLIPS but not in FLIPL cells. Additional data suggest that the PI3-kinase/Akt pathway is involved in constitutive NF-kB activation and differentially affected by FLIPL and FLIPS (Fig. 1D). Preliminary results in immunodeficient mice transplanted with transduced ML1 cells indicated the in vivo relevance of the differences between FLIPL and FLIPS with FLIPL cells engrafting earlier and showing earlier signs of sickness. Conclusions: FLIPL and FLIPS conferred resistance to TRAIL induced apoptosis but showed differential effects: Caspase-8/Bid was involved in the apoptosis pathway in FLIPS, but not in FLIPL cells. FLIPL cells’ resistance was due not only to caspase inhibition but to the recruitment of downstream anti-apoptotic pathways such as NF-kB and PI3K/Akt. In vivo data further substantiated the antiapoptotic/pro-survival behavior of FLIPL cells. Figure Figure

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