Knockdown of SIRT1 Suppresses Bladder Cancer Cell Proliferation and Migration and Induces Cell Cycle Arrest and Antioxidant Response through FOXO3a-Mediated Pathways

Bladder cancer (BCa) is one of the most common tumors, but its underlying mechanism has not been fully clarified. Our transcriptome analysis suggested a close link of Sirtuins, Peroxisome Proliferator-Activated Receptor (PPAR), cell cycle regulation, reactive oxygen species (ROS) metabolism, and Forkhead Box Class O (FOXO) signaling pathway in BCa. SIRT1 is a key member of Sirtuins, playing important roles in aging and energy metabolism, which has been reported to be involved in various metabolic diseases and tumors. We observed that SIRT1 was upregulated in BCa tissues at both mRNA and protein levels. By establishing a SIRT1-knockdown BCa cell model, our results suggested that proliferation and viability were suppressed. Moreover, migration rate was inhibited as well, possibly via reduction of epithelial-mesenchymal transition (EMT). In addition, cell cycle arrest was significantly induced, consisting with strongly decreased proteins involved (CDK2/4/6). Furthermore, ROS production was slightly reduced, accompanied by increasing of antioxidant enzymes and total/acetylated FOXO3a. Consistently with our Path-net analysis, we observed no significant alteration of apoptosis in the SIRT1-knockdown BCa cells. Taken together, our results suggested that SIRT1 deficiency in BCa cells could suppress cell viability by activating antioxidant response and inducing cell cycle arrest possibly via FOXO3a-related pathways.

[1]  Z. Meng,et al.  Decreased TRPM7 inhibits activities and induces apoptosis of bladder cancer cells via ERK1/2 pathway , 2016, Oncotarget.

[2]  E. Garrett-Mayer,et al.  Regulation of the Tumor Suppressor FOXO3 by the Thromboxane-A2 Receptors in Urothelial Cancer , 2014, PloS one.

[3]  Wei Gao,et al.  miR-543 promotes gastric cancer cell proliferation by targeting SIRT1. , 2016, Biochemical and biophysical research communications.

[4]  O. Leo,et al.  Sirtuin deacylases: a molecular link between metabolism and immunity , 2013, Journal of leukocyte biology.

[5]  A. Shen,et al.  PIASy mediates hypoxia-induced SIRT1 transcriptional repression and epithelial-to-mesenchymal transition in ovarian cancer cells , 2013, Journal of Cell Science.

[6]  D. Faller,et al.  SIRT1 enhances matrix metalloproteinase‐2 expression and tumor cell invasion in prostate cancer cells , 2013, The Prostate.

[7]  J. Buolamwini.  Cell cycle molecular targets in novel anticancer drug discovery. , 2000, Current pharmaceutical design.

[8]  De-Shuang Huang,et al.  Mining the bladder cancer-associated genes by an integrated strategy for the construction and analysis of differential co-expression networks , 2015, BMC Genomics.

[9]  Johannes Gerdes,et al.  The Ki‐67 protein: From the known and the unknown , 2000, Journal of cellular physiology.

[10]  Yingying Zhou,et al.  Over-expression of Sirt1 contributes to chemoresistance and indicates poor prognosis in serous epithelial ovarian cancer (EOC) , 2015, Medical Oncology.

[11]  D. Sinclair,et al.  Mammalian sirtuins: biological insights and disease relevance. , 2010, Annual review of pathology.

[12]  Wei Jiang,et al.  Simvastatin induces cell cycle arrest and inhibits proliferation of bladder cancer cells via PPARγ signalling pathway , 2016, Scientific Reports.

[13]  G. Demetri,et al.  Antiproliferative Effects of CDK4/6 Inhibition in CDK4-Amplified Human Liposarcoma In Vitro and In Vivo , 2014, Molecular Cancer Therapeutics.

[14]  Kaiyu Qian,et al.  Capsaicin Suppresses Cell Proliferation, Induces Cell Cycle Arrest and ROS Production in Bladder Cancer Cells through FOXO3a-Mediated Pathways , 2016, Molecules.

[15]  Tahir Ali Chohan,et al.  Cyclin-dependent kinase-2 as a target for cancer therapy: progress in the development of CDK2 inhibitors as anti-cancer agents. , 2014, Current medicinal chemistry.

[16]  Yilin Zhang,et al.  Microarray-based bioinformatics analysis of osteoblasts on TiO2 nanotube layers. , 2012, Colloids and surfaces. B, Biointerfaces.

[17]  T. Wada,et al.  Sirtuin1 expression predicts the efficacy of neoadjuvant chemotherapy for locally advanced uterine cervical cancer. , 2015, Molecular and clinical oncology.

[18]  M. Malumbres,et al.  Cyclin-dependent kinases , 2014, Genome Biology.

[19]  R. Frye,et al.  Characterization of five human cDNAs with homology to the yeast SIR2 gene: Sir2-like proteins (sirtuins) metabolize NAD and may have protein ADP-ribosyltransferase activity. , 1999, Biochemical and biophysical research communications.

[20]  R. Montironi,et al.  Metabolic phenotype of bladder cancer. , 2016, Cancer treatment reviews.

[21]  Shufeng Zhou,et al.  Pro-apoptotic and pro-autophagic effects of the Aurora kinase A inhibitor alisertib (MLN8237) on human osteosarcoma U-2 OS and MG-63 cells through the activation of mitochondria-mediated pathway and inhibition of p38 MAPK/PI3K/Akt/mTOR signaling pathway , 2015, Drug design, development and therapy.

[22]  A. Larue,et al.  Catalase inhibits ionizing radiation-induced apoptosis in hematopoietic stem and progenitor cells. , 2015, Stem cells and development.

[23]  Shufeng Zhou,et al.  Induction of apoptosis and autophagy via sirtuin1- and PI3K/Akt/mTOR-mediated pathways by plumbagin in human prostate cancer cells , 2015, Drug design, development and therapy.

[24]  L. Donini,et al.  Circulating SIRT1 inversely correlates with epicardial fat thickness in patients with obesity. , 2016, Nutrition, metabolism, and cardiovascular diseases : NMCD.

[25]  L. Guarente,et al.  Extrachromosomal rDNA Circles— A Cause of Aging in Yeast , 1997, Cell.

[26]  Jinsong Liu,et al.  Selective killing of oncogenically transformed cells through a ROS-mediated mechanism by beta-phenylethyl isothiocyanate. , 2006, Cancer cell.

[27]  W. Jia,et al.  Association of Genetic Variants of SIRT1 With Type 2 Diabetes Mellitus. , 2015, Gene expression.

[28]  Shreya Sharma,et al.  Expression/localization patterns of sirtuins (SIRT1, SIRT2, and SIRT7) during progression of cervical cancer and effects of sirtuin inhibitors on growth of cervical cancer cells , 2015, Tumor Biology.

[29]  Yul Ri Chung,et al.  Distinctive role of SIRT1 expression on tumor invasion and metastasis in breast cancer by molecular subtype. , 2015, Human pathology.

[30]  S. Shibata,et al.  Carnosic acid protects starvation-induced SH-SY5Y cell death through Erk1/2 and Akt pathways, autophagy, and FoxO3a , 2016, International journal of food sciences and nutrition.

[31]  B. Aggarwal,et al.  Upsides and downsides of reactive oxygen species for cancer: the roles of reactive oxygen species in tumorigenesis, prevention, and therapy. , 2012, Antioxidants & redox signaling.

[32]  C. Mathers,et al.  Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012 , 2015, International journal of cancer.

[33]  Eccentric localization of catalase to protect chromosomes from oxidative damages during meiotic maturation in mouse oocytes , 2016, Histochemistry and Cell Biology.

[34]  L. Kiemeney,et al.  Epidemiology and risk factors of urothelial bladder cancer. , 2013, European urology.

[35]  Junhong Wang,et al.  [Effect of metformin on the expression of SIRT1 and UCP2 in rat liver of type 2 diabetes mellitus and nonalcoholic fatty liver]. , 2013, Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences.

[36]  M. Cheng,et al.  Dietary Blueberry and Bifidobacteria Attenuate Nonalcoholic Fatty Liver Disease in Rats by Affecting SIRT1-Mediated Signaling Pathway , 2014, Oxidative medicine and cellular longevity.

[37]  Barry Halliwell,et al.  Oxidative stress and cancer: have we moved forward? , 2007, The Biochemical journal.

[38]  Y. Miyagi,et al.  SIRT1 expression is associated with a poor prognosis, whereas DBC1 is associated with favorable outcomes in gastric cancer , 2014, Cancer medicine.

[39]  Lihua Zhang,et al.  MiR-204 down regulates SIRT1 and reverts SIRT1-induced epithelial-mesenchymal transition, anoikis resistance and invasion in gastric cancer cells , 2013, BMC Cancer.

[40]  D. Newby,et al.  Effects of the small molecule SIRT1 activator, SRT2104 on arterial stiffness in otherwise healthy cigarette smokers and subjects with type 2 diabetes mellitus , 2016, Open Heart.

[41]  Friedrich-Carl von Rundstedt,et al.  Integrative Pathway Analysis of Metabolic Signature in Bladder Cancer: A Linkage to The Cancer Genome Atlas Project and Prediction of Survival , 2016, The Journal of urology.

[42]  Ji-min Cao,et al.  A novel crosstalk between BRCA1 and sirtuin 1 in ovarian cancer , 2014, Scientific Reports.

[43]  Minoru Kanehisa,et al.  KEGG as a reference resource for gene and protein annotation , 2015, Nucleic Acids Res..

[44]  D. Faller,et al.  SIRT1 induces EMT by cooperating with EMT transcription factors and enhances prostate cancer cell migration and metastasis , 2012, Oncogene.

[45]  Yi-Hui Lee,et al.  Capsaicin Inhibits Multiple Bladder Cancer Cell Phenotypes by Inhibiting Tumor-Associated NADH Oxidase (tNOX) and Sirtuin1 (SIRT1) , 2016, Molecules.

[46]  Jianping Ye,et al.  Diet‐induced obesity and insulin resistance are associated with brown fat degeneration in SIRT1‐deficient mice , 2016, Obesity.

[47]  E. Şenateş,et al.  SIRT1 as a potential therapeutic target for treatment of nonalcoholic fatty liver disease , 2011, Medical science monitor : international medical journal of experimental and clinical research.

[48]  J. Sweatt,et al.  Obesity Weighs down Memory through a Mechanism Involving the Neuroepigenetic Dysregulation of Sirt1 , 2016, The Journal of Neuroscience.

[49]  Brad T. Sherman,et al.  DAVID: Database for Annotation, Visualization, and Integrated Discovery , 2003, Genome Biology.

[50]  Haitao Wang,et al.  Involvement of PI3K/Akt/FoxO3a and PKA/CREB Signaling Pathways in the Protective Effect of Fluoxetine Against Corticosterone-Induced Cytotoxicity in PC12 Cells , 2016, Journal of Molecular Neuroscience.

[51]  中尾 光輝,et al.  KEGG(Kyoto Encyclopedia of Genes and Genomes)〔和文〕 (特集 ゲノム医学の現在と未来--基礎と臨床) -- (データベース) , 2000 .

[52]  N. Z. Zur Nieden,et al.  Glucose-Induced Oxidative Stress Reduces Proliferation in Embryonic Stem Cells via FOXO3A/β-Catenin-Dependent Transcription of p21cip1 , 2016, Stem cell reports.

[53]  M. Cascante,et al.  Cyclin-dependent kinases 4 and 6 control tumor progression and direct glucose oxidation in the pentose cycle , 2012, Metabolomics.

[54]  M. Pellegrino,et al.  SIRT1 is involved in oncogenic signaling mediated by GPER in breast cancer , 2015, Cell Death and Disease.

[55]  O. Shaker,et al.  Association between SIRT1 Gene Polymorphisms and Breast Cancer in Egyptians , 2016, PloS one.

[56]  Lijia Xiao,et al.  Orphan nuclear receptor TLX functions as a potent suppressor of oncogene‐induced senescence in prostate cancer via its transcriptional co‐regulation of the CDKN1A (p21WAF1/CIP1) and SIRT1 genes , 2015, The Journal of pathology.

[57]  R. Frye,et al.  Phylogenetic classification of prokaryotic and eukaryotic Sir2-like proteins. , 2000, Biochemical and biophysical research communications.

[58]  M. McVey,et al.  The SIR2/3/4 complex and SIR2 alone promote longevity in Saccharomyces cerevisiae by two different mechanisms. , 1999, Genes & development.

[59]  J. Milner,et al.  Oncogenic viral protein HPV E7 up-regulates the SIRT1 longevity protein in human cervical cancer cells , 2009, Aging.

[60]  R. Frazzi,et al.  Resveratrol‐mediated apoptosis of hodgkin lymphoma cells involves SIRT1 inhibition and FOXO3a hyperacetylation , 2013, International journal of cancer.