Hesperidin induces apoptosis and triggers autophagic markers through inhibition of Aurora-A mediated phosphoinositide-3-kinase/Akt/mammalian target of.

Slides:



Advertisements
Similar presentations
Iron citrate reduces high phosphate-induced vascular calcification by inhibiting apoptosis  Paola Ciceri, Francesca Elli, Paola Braidotti, Monica Falleni,
Advertisements

Inhibition of autophagy augments 5-fluorouracil chemotherapy in human colon cancer in vitro and in vivo model  Jie Li, Ni Hou, Ahmad Faried, Soichi Tsutsumi,
Darren M Brown, Erkki Ruoslahti  Cancer Cell 
Δ-Tocotrienol treatment is more effective against hypoxic tumor cells than normoxic cells: potential implications for cancer therapy  Akira Shibata, Kiyotaka.
The role of galectin-1 in in vitro and in vivo photodynamic therapy with a galactodendritic porphyrin  Patrícia M.R. Pereira, Sandrina Silva, José S.
Anandamide inhibits the Wnt/β-catenin signalling pathway in human breast cancer MDA MB 231 cells  Chiara Laezza, Alba D’Alessandro, Simona Paladino, Anna.
Expression of suppressor of cytokine signaling 1 and 3 in ligature-induced periodontitis in rats  João Antonio Chaves de Souza, Andressa Vilas Boas Nogueira,
Volume 15, Pages (February 2017)
Vascular endothelial growth factor-C derived from CD11b+ cells induces therapeutic improvements in a murine model of hind limb ischemia  Go Kuwahara,
Selective interference of mTORC1/RAPTOR protects against human disc cellular apoptosis, senescence, and extracellular matrix catabolism with Akt and autophagy.
Amanda M. Nelson, Kathryn L. Gilliland, Zhaoyuan Cong, Diane M
Traditional extract of Pithecellobium dulce fruits protects mice against CCl4 induced renal oxidative impairments and necrotic cell death  Pabitra Bikash.
Canonical Wnt/β-catenin signaling mediates transforming growth factor-β1-driven podocyte injury and proteinuria  Dan Wang, Chunsun Dai, Yingjian Li, Youhua.
T. Minashima, K.A. Campbell, S.R. Hadley, Y. Zhang, T. Kirsch 
KIND1 Loss Sensitizes Keratinocytes to UV-Induced Inflammatory Response and DNA Damage  Xiaoling Zhang, Suju Luo, Joseph Wu, Long Zhang, Wen-hui Wang,
Expression of KRAS in the endometrium of early pregnant mice and its effect during embryo implantation  Xia Long, Min Zhang, Xuemei Chen, Junlin He, Yubin.
Volume 43, Issue 6, Pages (December 2015)
Hyperphosphatemia induces protective autophagy in endothelial cells through the inhibition of Akt/mTOR signaling  Yu-Juei Hsu, MD, PhD, Shih-Che Hsu,
Modification of Alternative Splicing of Mcl-1 Pre-mRNA Using Antisense Morpholino Oligonucleotides Induces Apoptosis in Basal Cell Carcinoma Cells  Jeng-Jer.
Volume 69, Issue 4, Pages (February 2006)
Volume 117, Issue 3, Pages (September 1999)
Hyperphosphatemia induces protective autophagy in endothelial cells through the inhibition of Akt/mTOR signaling  Yu-Juei Hsu, MD, PhD, Shih-Che Hsu,
Volume 138, Issue 1, Pages (January 2010)
Decreased Expression of Caveolin-1 Contributes to the Pathogenesis of Psoriasiform Dermatitis in Mice  Yukie Yamaguchi, Yuko Watanabe, Tomoya Watanabe,
Hepatoprotective properties of kombucha tea against TBHP-induced oxidative stress via suppression of mitochondria dependent apoptosis  Semantee Bhattacharya,
Volume 62, Issue 4, Pages (October 2002)
Dienogest enhances autophagy induction in endometriotic cells by impairing activation of AKT, ERK1/2, and mTOR  JongYeob Choi, Ph.D., MinWha Jo, M.S.,
Ropivacaine- and bupivacaine-induced death of rabbit annulus fibrosus cells in vitro: involvement of the mitochondrial apoptotic pathway  X.-Y. Cai, Y.
Volume 88, Issue 3, Pages (September 2015)
J.R. Liu, C. Baek, X.H. Han, P. Shoureshi, S.G. Soriano 
Selective interference of mTORC1/RAPTOR protects against human disc cellular apoptosis, senescence, and extracellular matrix catabolism with Akt and autophagy.
Inhibition of complement C5 reduces local and remote organ injury after intestinal ischemia/reperfusion in the rat  Koichiro Wada, Michael C. Montalto,
Combining the Multitargeted Tyrosine Kinase Inhibitor Vandetanib with the Antiestrogen Fulvestrant Enhances Its Antitumor Effect in Non-small Cell Lung.
CaMKII inhibition in human primary and pluripotent stem cell-derived chondrocytes modulates effects of TGFβ and BMP through SMAD signaling  B. Saitta,
Volume 73, Issue 5, Pages (March 2008)
Mechanisms of the proteinuria induced by Rho GTPases
Volume 67, Issue 4, Pages (April 2005)
Volume 134, Issue 3, Pages (March 2008)
Volume 69, Issue 2, Pages (August 2018)
Nitric oxide from rat liver sinusoidal endothelial cells induces apoptosis in IFN γ- sensitized CC531s colon carcinoma cells  Katrien Vekemans, Filip Braet,
Enhanced expression of glucose transporter-1 in vascular smooth muscle cells via the Akt/tuberous sclerosis complex subunit 2 (TSC2)/mammalian target.
Role of vitamin C and selenium in attenuation of nicotine induced oxidative stress, P53 and Bcl2 expression in adult rat spleen  Marwa A. Ahmed, K.H.
SOX4 Promotes Proliferative Signals by Regulating Glycolysis through AKT Activation in Melanoma Cells  Wei Dai, Xinyuan Xu, Shuli Li, Jingjing Ma, Qiong.
FcɛRI cross-linking and IL-3 protect human basophils from intrinsic apoptotic stress  Lionel Rohner, MSc, Ramona Reinhart, PhD, Björn Hagmann, PhD, Andrea.
Volume 7, Issue 2, Pages (February 2010)
Volume 81, Issue 5, Pages (March 2012)
Volume 139, Issue 3, Pages e6 (September 2010)
IL-27 Suppresses Antimicrobial Activity in Human Leprosy
Volume 79, Issue 9, Pages (May 2011)
Darren M Brown, Erkki Ruoslahti  Cancer Cell 
GLI2 Is a Regulator of β-Catenin and Is Associated with Loss of E-Cadherin, Cell Invasiveness, and Long-Term Epidermal Regeneration  Eleni Pantazi, Emilios.
Molecular Therapy - Methods & Clinical Development
Noritaka Oyama, Keiji Iwatsuki, Yoshimi Homma, Fumio Kaneko 
Volume 92, Issue 3, Pages (September 2017)
J.P O'Rourke, H Hiraragi, K Urban, M Patel, J.C Olsen, B.A Bunnell 
Inducible Nitric Oxide Synthase Up-Regulates Notch-1 in Mouse Cholangiocytes: Implications for Carcinogenesis  Norihisa Ishimura, Steven F. Bronk, Gregory.
The IL-6 Trans-Signaling-STAT3 Pathway Mediates ECM and Cellular Proliferation in Fibroblasts from Hypertrophic Scar  Sutapa Ray, Xiaoxi Ju, Hong Sun,
Hydroxychloroquine Modulates Metabolic Activity and Proliferation and Induces Autophagic Cell Death of Human Dermal Fibroblasts  Bettina Ramser, Agatha.
Xin Xie, Tomas Venit, Nizar Drou, Piergiorgio Percipalle
EVA1A/TMEM166 Regulates Embryonic Neurogenesis by Autophagy
Volume 64, Issue 6, Pages (December 2003)
Volume 41, Issue 5, Pages (November 2004)
Expression of the peripheral-type benzodiazepine receptor and apoptosis induction in hepatic stellate cells  Richard Fischer, Marcus Schmitt, Johannes.
Volume 90, Issue 3, Pages (May 2016)
Lin Mu, Ph. D. , Wei Zheng, Ph. D. , M. D. , Liang Wang, Ph. D
Fang Du, Qing Yu, Allen Chen, Doris Chen, Shirley ShiDu Yan 
Fig. 1. DEL-1 is expressed by human and mouse osteoclasts.
Mathematical Modeling Highlights the Complex Role of AKT in TRAIL-Induced Apoptosis of Colorectal Carcinoma Cells  Matthew W. Anderson, Joanna J. Moss,
Fig. 2. Col IV–Ac2-26 NPs increase subendothelial collagen in Ldlr−/− mice with established atherosclerosis. Col IV–Ac2-26 NPs increase subendothelial.
Identification of Kinases Responsible for p53-Dependent Autophagy
Presentation transcript:

Hesperidin induces apoptosis and triggers autophagic markers through inhibition of Aurora-A mediated phosphoinositide-3-kinase/Akt/mammalian target of rapamycin and glycogen synthase kinase-3 beta signalling cascades in experimental colon carcinogenesis  Gowrikumar Saiprasad, Palanivel Chitra, Ramar Manikandan, Ganapasam Sudhandiran  European Journal of Cancer  Volume 50, Issue 14, Pages 2489-2507 (September 2014) DOI: 10.1016/j.ejca.2014.06.013 Copyright © 2014 Elsevier Ltd Terms and Conditions

Fig. 1 (A) Schematic diagram of the experimental protocol. Mice received AOM once a week for three consecutive weeks for inducing colon cancer. Hesperidin was given orally starting a week before AOM injection and continued till the last AOM injection (initiation) or were administered hesperidin one week after the end of the last AOM injection and continued till the end of experiment (post-initiation). (B) The levels of CEA (carcino embroyonic antigen) were estimated in serum of control and experimental groups of animals. Each value is expressed as mean±S.D. of three separate experiments (n=3). Hypothesis testing method included a one-way analysis of variance (ANOVA) followed by the post hoc Tukey’s test. Results are statistically significance at P<0.05. Values sharing following symbols differ significantly, ∗ – versus control; # – versus AOM; † – versus AOM+HES (post-initiation). No significant difference was observed between control and hesperidin alone administered group of animals (AOM – Azoxymethane, HES – Hesperidin). European Journal of Cancer 2014 50, 2489-2507DOI: (10.1016/j.ejca.2014.06.013) Copyright © 2014 Elsevier Ltd Terms and Conditions

Fig. 2 (A) Representative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of transcripts for Aurora-A in the colonic tissues of control and experimental groups of animal. GAPDH (glyceraldehyde 3-phosphate dehydrogenase) served as internal control. Lane 1 – Control, Lane 2 – AOM, Lane 3 – HES+AOM (initiation), Lane 4 – AOM+HES (post-initiation), Lane 5 – HES. (B) The quantitative data represent the band intensity levels assessed using densitometry. (C) Immunoblots of Aurora-A in the colonic tissues of control and experimental groups of animal. β-Actin served as internal control. (D) The quantitative data represent the corresponding protein levels assessed using densitometry. Y-axis represents relative intensity (arbitrary units)/gene expression (fold change). Each value is expressed as mean±S.D. of three independently performed experiments (n=3). Hypothesis testing method included a one-way analysis of variance (ANOVA) followed by the post hoc Tukey’s test. Results are statistically significant at P<0.05. Values sharing the following symbols differ significantly, ∗ – versus control; # – versus AOM; † – versus AOM+HES (post-initiation). No significant difference was observed between control and hesperidin alone administered group of animals (AOM – Azoxymethane, HES – Hesperidin). European Journal of Cancer 2014 50, 2489-2507DOI: (10.1016/j.ejca.2014.06.013) Copyright © 2014 Elsevier Ltd Terms and Conditions

Fig. 3 Representative photographs for the immunohistochemical staining of Aurora-A, 20× (Scale bar – 50μm). (A) Colon tissue sections of control animals (control); the square with enlarged image highlights scarce expression of Aurora-A. (B) Colon tissue sections of AOM induced animals (AOM); the square with enlarged image highlights the significantly increased expression of Aurora-A. Colonic tissue sections of hesperidin treated animals at both initiation (HES+AOM) (C) as well as post-initiation (AOM+HES) phase (D); the square with enlarged image highlights the markedly decreased immunoreactivity of Aurora-A. (E) Colon sections of hesperidin alone administered animals (HES) showing scarce expression of Aurora-A similar to control. (F) The Aurora-A expression was quantified by averaging positive cells across 20 randomly selected fields. Each value is expressed as mean±S.D. of three separate experiments (n=3). Hypothesis testing method included a one-way analysis of variance (ANOVA) followed by the post hoc Tukey’s test. Results are statistically significance at P<0.05. Values sharing following symbols differ significantly, ∗ – versus control; # – versus AOM; † – versus AOM+HES (post-initiation). No significant difference was observed between control and hesperidin alone administered group of animals (AOM – Azoxymethane, HES – Hesperidin). European Journal of Cancer 2014 50, 2489-2507DOI: (10.1016/j.ejca.2014.06.013) Copyright © 2014 Elsevier Ltd Terms and Conditions

Fig. 4 (A) Representative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of transcripts for Bcl-2 and Bax in the colonic tissues of control and experimental groups of animals. GAPDH (glyceraldehyde 3-phosphate dehydrogenase) served as internal control. Lane 1 – Control, Lane 2 – AOM, Lane 3 – HES+AOM (initiation), Lane 4 – AOM+HES (post-initiation), Lane 5 – HES. (B and C) The quantitative data represent the corresponding levels of mRNA transcripts assessed using densitometry. (D) Immunoblotting data of mitochondrial expressions of Bcl-2, Bax, cytochrome c in the colonic tissues of control and experimental groups of animals. (E) Immunoblotting data of cytoplasmic expressions of Bax, cytochrome c and caspase-3 and 9 in the colonic tissues of control and experimental groups of animals. VDAC1 served as internal control for mitochondrial protein analysis and β-Actin served as internal control for the cytoplasmic protein analysis. (F and G) The quantitative data represent the corresponding protein levels assessed using densitometry. (H) The graphical data represent the ratio between Bax/Bcl-2 (protein expression) assessed using densitometry. (I) DNA fragmentation analysis of the corresponding genomic DNA of control and other experimental group of animals. Y-axis represents relative intensity (arbitrary units)/gene expression (fold change). Each value is expressed as mean±S.D. of three independently performed experiments (n=3). Hypothesis testing method included a one-way analysis of variance (ANOVA) followed by the post hoc Tukey’s test. Results are statistically significant at P<0.05. Values sharing the following symbols differ significantly, ∗ – versus control; # – versus AOM; † – versus AOM+HES (post-initiation). No significant difference was observed between control and hesperidin alone administered group of animals (AOM – Azoxymethane, HES – Hesperidin). European Journal of Cancer 2014 50, 2489-2507DOI: (10.1016/j.ejca.2014.06.013) Copyright © 2014 Elsevier Ltd Terms and Conditions

Fig. 5 (A) Representative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of transcripts for p53 and p21 in the colonic tissues of control and experimental groups of animals. GAPDH (glyceraldehyde 3-phosphate dehydrogenase) served as internal control. Lane 1 – Control, Lane 2 – AOM, Lane 3 – HES+AOM (initiation), Lane 4 – AOM+HES (post-initiation), Lane 5 – HES. (B) The quantitative data represent the corresponding levels of mRNA transcripts assessed using densitometry. (C) Immunoblotting data of p53 and p21 in the colonic tissues of control and experimental groups of animals. β-Actin served as internal control. (D) The quantitative data represent the corresponding protein levels assessed using densitometry. Y-axis represents relative intensity (arbitrary units)/gene expression (fold change). Each value is expressed as mean±S.D. of three independently performed experiments (n=3). Hypothesis testing method included a one-way analysis of variance (ANOVA) followed by the post hoc Tukey’s test. Results are statistically significance at P<0.05. Values sharing following symbols differ significantly, ∗ – versus control; # – versus AOM; † – versus AOM+HES (post-initiation). No significant difference was observed between control and hesperidin alone administered group of animals (AOM – Azoxymethane, HES – Hesperidin). European Journal of Cancer 2014 50, 2489-2507DOI: (10.1016/j.ejca.2014.06.013) Copyright © 2014 Elsevier Ltd Terms and Conditions

Fig. 6 (A) The immunoblotting data of p-PI3K, p-AktSer-473, p-Akt Thr-308, total Akt and phosphatase and tensin homologue (PTEN) from the colon tissues of control and experimental groups of animals. β-Actin served as internal control. Lane 1 – Control, Lane 2 – AOM, Lane 3 – HES+AOM (initiation), Lane 4 – AOM+HES (post-initiation), Lane 5 – HES. (B) The quantitative data represent the corresponding protein levels assessed using densitometry. (C) The graphical data represent the ratio between p-Akt/total Akt assessed using densitometry. Y-axis represents relative intensity (arbitrary units)/gene expression (fold change). Each value is expressed as mean±S.D. of three independently performed experiments (n=3). Hypothesis testing method included a one-way analysis of variance (ANOVA) followed by the post hoc Tukey’s test. Results are statistically significance at P<0.05. Values sharing following symbols differ significantly, ∗ – versus control; # – versus AOM; † – versus AOM+HES (post-initiation). No significant difference was observed between control and hesperidin alone administered group of animals (AOM – Azoxymethane, HES – Hesperidin). European Journal of Cancer 2014 50, 2489-2507DOI: (10.1016/j.ejca.2014.06.013) Copyright © 2014 Elsevier Ltd Terms and Conditions

Fig. 7 Confocal microscopic analysis of p-PI3K expression in the colonic sections of control and experimental groups of animals. Images shown are representative of three separate experiments (n=3). Tissue sections were immunostained with anti-p-PI3K antibody and FITC conjugated secondary antibody (green). Tissue sections were counter stained using PI (red) to stain nucleus (Scale bar – 50μm). Slides were visualised under the confocal microscope (Leica TCS-SP2 XL) using excitation/emission wavelength for PI – 529nm/620nm and FITC – 494nm/525nm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) European Journal of Cancer 2014 50, 2489-2507DOI: (10.1016/j.ejca.2014.06.013) Copyright © 2014 Elsevier Ltd Terms and Conditions

Fig. 8 (A) Representative immunoblotting data for p-mTOR, Beclin-1 and LC3-II from the colonic tissues of control and experimental groups of animals. β-Actin served as internal control. Lane 1 – Control, Lane 2 – AOM, Lane 3 – HES+AOM (initiation), Lane 4 – AOM+HES (post-initiation), Lane 5 – HES. (B) The quantitative data represent the corresponding protein levels assessed using densitometry. (C) Representative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of transcripts for Beclin-1 and LC3 in the colonic tissues of control and experimental groups of animals. GAPDH (glyceraldehyde 3-phosphate dehydrogenase) served as internal control. (D) The quantitative data represent the corresponding levels of mRNA transcripts assessed using densitometry. Y-axis represents relative intensity (arbitrary units)/gene expression (fold change). Each value is expressed as mean±S.D. of three independently performed experiments (n=3). Hypothesis testing method included a one-way analysis of variance (ANOVA) followed by post hoc Tukey’s test. Results are statistically significance at P<0.05. Values sharing following symbols differ significantly, ∗ – versus control; # – versus AOM; † – versus AOM+HES (post-initiation). No significant difference was observed between control and hesperidin alone administered group of animals (AOM – Azoxymethane, HES – Hesperidin). European Journal of Cancer 2014 50, 2489-2507DOI: (10.1016/j.ejca.2014.06.013) Copyright © 2014 Elsevier Ltd Terms and Conditions

Fig. 9 (A) The immunoblotting data of p-GSK-3βSer-9, Cyclin-D1 and β-catenin from the colon tissues of control and experimental groups of animals. β-Actin served as internal control for the cytoplasmic protein and Histone H2B served as internal control for the nuclear protein analysis. Lane 1 – Control, Lane 2 – AOM, Lane 3 – HES+AOM (initiation), Lane 4 – AOM+HES (post-initiation), Lane 5 – HES. (B) The quantitative data represent the corresponding protein levels assessed using densitometry. (C) Representative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of transcripts for c-myc and c-jun in the colonic tissues of control and experimental groups of animals. GAPDH (glyceraldehyde 3-phosphate dehydrogenase) served as internal control. (D) The quantitative data represent the corresponding levels of mRNA transcripts assessed using densitometry. Y-axis represents relative intensity (arbitrary units)/gene expression (fold change). Each value is expressed as mean±S.D. of three independently performed experiments (n=3). Hypothesis testing method included a one-way analysis of variance (ANOVA) followed by the post hoc Tukey’s test. Results are statistically significance at P<0.05. Values sharing following symbols differ significantly, ∗ – versus control; # – versus AOM; † – versus AOM+HES (post-initiation). No significant difference was observed between control and HES alone administered group of animals (AOM – Azoxymethane, HES – Hesperidin). European Journal of Cancer 2014 50, 2489-2507DOI: (10.1016/j.ejca.2014.06.013) Copyright © 2014 Elsevier Ltd Terms and Conditions

Fig. 10 Representative scanning electron microscopic images of the colonic tissues of control and experimental groups of animals. (A, H) Control, H - enlarged image of A, (B–D) AOM induced animals, (E, G) AOM induced animals treated with hesperidin at initiation phase (HES+AOM), G - enlarged image of (E, F) AOM induced animals treated with hesperidin at post-initiation phase (AOM+HES) (AOM – Azoxymethane, HES – hesperidin). European Journal of Cancer 2014 50, 2489-2507DOI: (10.1016/j.ejca.2014.06.013) Copyright © 2014 Elsevier Ltd Terms and Conditions