22 results on '"Cinghu, S."'
Search Results
2. 128 FTS (Fused Toes Homolog) silencing increases the effect of cisplatin through inhibiting EGFR-mediated repair of DNA damage in cervix cancer cells
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Park, W.Y., primary, Ds, P., additional, Cinghu, S., additional, and Yu, J.R., additional
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- 2015
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3. 149: Nuclear translocation of FTS (Fused Toes Homolog) is required for EGFR phosphorylation and confers radiation resistance on uterine cervix cancer
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Park, W.-Y., primary, Anandharaj, A., additional, Cinghu, S., additional, and Yu, J.-R., additional
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- 2014
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4. 8045 POSTER Fused Toes Homolog is a Novel Oncoprotein Involved in Uterine Cervical Carcinogenesis and a Potential Diagnostic Marker for Cervical Cancer
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Cinghu, S., primary, Anandharaj, A., additional, Lee, H., additional, Yu, J., additional, and Park, W., additional
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- 2011
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5. Nucleolin maintains embryonic stem cell homeostasis by shielding against differentiation-inducing redox-imbalance induced oxidative stress
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Cinghu, S., Yellaboina, S., Freudenberg, J. M., Ghosh, S., Zheng, X., Oldfield, A. J., Lackford, B. L., Dmitri Zaykin, Hu, G., and Jothi, R.
6. Decoding the function of bivalent chromatin in development and cancer.
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Kumar D, Cinghu S, Oldfield AJ, Yang P, and Jothi R
- Abstract
Bivalent chromatin is characterized by the simultaneous presence of H3K4me3 and H3K27me3, histone modifications generally associated with transcriptionally active and repressed chromatin, respectively. Prevalent in embryonic stem cells (ESCs), bivalency is postulated to poise/prime lineage-controlling developmental genes for rapid activation during embryogenesis while maintaining a transcriptionally repressed state in the absence of activation cues; however, this hypothesis remains to be directly tested. Most gene promoters DNA hypermethylated in adult human cancers are bivalently marked in ESCs, and it was speculated that bivalency predisposes them for aberrant de novo DNA methylation and irreversible silencing in cancer, but evidence supporting this model is largely lacking. Here, we show that bivalent chromatin does not poise genes for rapid activation but protects promoters from de novo DNA methylation. Genome-wide studies in differentiating ESCs reveal that activation of bivalent genes is no more rapid than that of other transcriptionally silent genes, challenging the premise that H3K4me3 is instructive for transcription. H3K4me3 at bivalent promoters-a product of the underlying DNA sequence-persists in nearly all cell types irrespective of gene expression and confers protection from de novo DNA methylation. Bivalent genes in ESCs that are frequent targets of aberrant hypermethylation in cancer are particularly strongly associated with loss of H3K4me3/bivalency in cancer. Altogether, our findings suggest that bivalency protects reversibly repressed genes from irreversible silencing and that loss of H3K4me3 may make them more susceptible to aberrant DNA methylation in diseases such as cancer. Bivalency may thus represent a distinct regulatory mechanism for maintaining epigenetic plasticity., (Published by Cold Spring Harbor Laboratory Press.)
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- 2021
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7. Transcriptional Regulation of Structural and Functional Adaptations in a Developing Adulthood Myocardium.
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Sunny S, Challa AK, Qiao A, Jyothidasan A, Krishnamurthy P, Ramamurthy MT, Crossman DK, Pogwizd S, Cinghu S, and Rajasekaran NS
- Abstract
The development of the heart follows a synergic action of several signaling pathways during gestational, pre- & postnatal stages. The current study aimed to investigate whether the myocardium experiences transcriptional changes during the transition from post-natal to adult hood stages. Herein, we used C57/B16/J mice at 4 (28- days; post-natal/PN) and 20 weeks (adulthood/AH) of ages and employed the next generation RNAseq (NGS) to profile the transcriptome and echocardiography analysis to monitor the structural/functional changes in the heart. NGS-based RNA-seq revealed that 1215 genes were significantly upregulated and 2549 were down regulated in the AH versus PN hearts, indicating a significant transcriptional change during this transition. A synchronized cardiac transcriptional regulation through cell cycle, growth hormones, redox homeostasis and metabolic pathways was noticed in both PN and AH hearts. Echocardiography reveals significant structural and functional (i.e. systolic/diastolic) changes during the transition of PN to adult stage. Particularly, a progressive decline in ejection fraction and cardiac output was observed in AH hearts. These structural adaptations are in line with critical signaling pathways that drive the maturation of heart during AH. Overall, we have presented a comprehensive transcriptomic analysis along with structural-functional relationship during the myocardial development in adult mice., Competing Interests: Competing Interests The authors have no competing interests to declare.
- Published
- 2021
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8. Transcriptional network dynamics during the progression of pluripotency revealed by integrative statistical learning.
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Kim HJ, Osteil P, Humphrey SJ, Cinghu S, Oldfield AJ, Patrick E, Wilkie EE, Peng G, Suo S, Jothi R, Tam PPL, and Yang P
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- Animals, Binding Sites genetics, Cell Differentiation genetics, Chromatin genetics, Gene Expression Regulation, Developmental genetics, Gene Regulatory Networks genetics, Genome genetics, Mice, Embryonic Development genetics, Mouse Embryonic Stem Cells metabolism, Pluripotent Stem Cells metabolism, Transcription Factors genetics
- Abstract
The developmental potential of cells, termed pluripotency, is highly dynamic and progresses through a continuum of naive, formative and primed states. Pluripotency progression of mouse embryonic stem cells (ESCs) from naive to formative and primed state is governed by transcription factors (TFs) and their target genes. Genomic techniques have uncovered a multitude of TF binding sites in ESCs, yet a major challenge lies in identifying target genes from functional binding sites and reconstructing dynamic transcriptional networks underlying pluripotency progression. Here, we integrated time-resolved 'trans-omic' datasets together with TF binding profiles and chromatin conformation data to identify target genes of a panel of TFs. Our analyses revealed that naive TF target genes are more likely to be TFs themselves than those of formative TFs, suggesting denser hierarchies among naive TFs. We also discovered that formative TF target genes are marked by permissive epigenomic signatures in the naive state, indicating that they are poised for expression prior to the initiation of pluripotency transition to the formative state. Finally, our reconstructed transcriptional networks pinpointed the precise timing from naive to formative pluripotency progression and enabled the spatiotemporal mapping of differentiating ESCs to their in vivo counterparts in developing embryos., (© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.)
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- 2020
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9. NF-Y controls fidelity of transcription initiation at gene promoters through maintenance of the nucleosome-depleted region.
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Oldfield AJ, Henriques T, Kumar D, Burkholder AB, Cinghu S, Paulet D, Bennett BD, Yang P, Scruggs BS, Lavender CA, Rivals E, Adelman K, and Jothi R
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- Animals, CCAAT-Binding Factor genetics, Cell Line, Chromatin genetics, Chromatin metabolism, Embryonic Stem Cells, Gene Knockdown Techniques, Mice, Nucleosomes genetics, Promoter Regions, Genetic genetics, RNA, Small Interfering metabolism, CCAAT-Binding Factor metabolism, Nucleosomes metabolism, Transcription Initiation Site, Transcription Initiation, Genetic
- Abstract
Faithful transcription initiation is critical for accurate gene expression, yet the mechanisms underlying specific transcription start site (TSS) selection in mammals remain unclear. Here, we show that the histone-fold domain protein NF-Y, a ubiquitously expressed transcription factor, controls the fidelity of transcription initiation at gene promoters in mouse embryonic stem cells. We report that NF-Y maintains the region upstream of TSSs in a nucleosome-depleted state while simultaneously protecting this accessible region against aberrant and/or ectopic transcription initiation. We find that loss of NF-Y binding in mammalian cells disrupts the promoter chromatin landscape, leading to nucleosomal encroachment over the canonical TSS. Importantly, this chromatin rearrangement is accompanied by upstream relocation of the transcription pre-initiation complex and ectopic transcription initiation. Further, this phenomenon generates aberrant extended transcripts that undergo translation, disrupting gene expression profiles. These results suggest NF-Y is a central player in TSS selection in metazoans and highlight the deleterious consequences of inaccurate transcription initiation.
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- 2019
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10. Multi-omic Profiling Reveals Dynamics of the Phased Progression of Pluripotency.
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Yang P, Humphrey SJ, Cinghu S, Pathania R, Oldfield AJ, Kumar D, Perera D, Yang JYH, James DE, Mann M, and Jothi R
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- Animals, Cell Differentiation physiology, Cell Lineage, Embryonic Stem Cells cytology, Epigenome genetics, Gene Expression Regulation, Developmental, Germ Layers cytology, Germ Layers metabolism, Humans, Proteome metabolism, Signal Transduction, Transcriptome genetics, Pluripotent Stem Cells metabolism, Pluripotent Stem Cells physiology
- Abstract
Pluripotency is highly dynamic and progresses through a continuum of pluripotent stem cell states. The two states that bookend the pluripotency continuum, naive and primed, are well characterized, but our understanding of the intermediate states and transitions between them remains incomplete. Here, we dissect the dynamics of pluripotent state transitions underlying pre- to post-implantation epiblast differentiation. Through comprehensive mapping of the proteome, phosphoproteome, transcriptome, and epigenome of embryonic stem cells transitioning from naive to primed pluripotency, we find that rapid, acute, and widespread changes to the phosphoproteome precede ordered changes to the epigenome, transcriptome, and proteome. Reconstruction of the kinase-substrate networks reveals signaling cascades, dynamics, and crosstalk. Distinct waves of global proteomic changes mark discrete phases of pluripotency, with cell-state-specific surface markers tracking pluripotent state transitions. Our data provide new insights into multi-layered control of the phased progression of pluripotency and a foundation for modeling mechanisms regulating pluripotent state transitions (www.stemcellatlas.org)., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)
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- 2019
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11. Intragenic Enhancers Attenuate Host Gene Expression.
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Cinghu S, Yang P, Kosak JP, Conway AE, Kumar D, Oldfield AJ, Adelman K, and Jothi R
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- Animals, CRISPR-Cas Systems, Cell Line, Chromatin chemistry, Chromatin metabolism, Embryoid Bodies cytology, Embryoid Bodies metabolism, Gene Editing, Mice, Mouse Embryonic Stem Cells cytology, Promoter Regions, Genetic, RNA genetics, RNA metabolism, RNA Polymerase II metabolism, Enhancer Elements, Genetic, Gene Expression Regulation, Mouse Embryonic Stem Cells metabolism, RNA Polymerase II genetics, Transcription Elongation, Genetic
- Abstract
Eukaryotic gene transcription is regulated at many steps, including RNA polymerase II (Pol II) recruitment, transcription initiation, promoter-proximal Pol II pause release, and transcription termination; however, mechanisms regulating transcription during productive elongation remain poorly understood. Enhancers, which activate gene transcription, themselves undergo Pol II-mediated transcription, but our understanding of enhancer transcription and enhancer RNAs (eRNAs) remains incomplete. Here we show that transcription at intragenic enhancers interferes with and attenuates host gene transcription during productive elongation. While the extent of attenuation correlates positively with nascent eRNA expression, the act of intragenic enhancer transcription alone, but not eRNAs, explains the attenuation. Through CRISPR/Cas9-mediated deletions, we demonstrate a physiological role for intragenic enhancer-mediated transcription attenuation in cell fate determination. We propose that intragenic enhancers not only enhance transcription of one or more genes from a distance but also fine-tune transcription of their host gene through transcription interference, facilitating differential utilization of the same regulatory element for disparate functions., (Published by Elsevier Inc.)
- Published
- 2017
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12. DNMT1 is essential for mammary and cancer stem cell maintenance and tumorigenesis.
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Pathania R, Ramachandran S, Elangovan S, Padia R, Yang P, Cinghu S, Veeranan-Karmegam R, Arjunan P, Gnana-Prakasam JP, Sadanand F, Pei L, Chang CS, Choi JH, Shi H, Manicassamy S, Prasad PD, Sharma S, Ganapathy V, Jothi R, and Thangaraju M
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- Animals, Blotting, Western, Breast Neoplasms metabolism, Cell Line, Cell Line, Tumor, DNA (Cytosine-5-)-Methyltransferase 1, DNA (Cytosine-5-)-Methyltransferases metabolism, DNA Methylation, Down-Regulation, Female, Humans, LIM-Homeodomain Proteins metabolism, MCF-7 Cells, Mammary Glands, Animal cytology, Mammary Glands, Animal growth & development, Mammary Neoplasms, Experimental metabolism, Mice, Microscopy, Fluorescence, Neoplastic Stem Cells cytology, Stem Cells metabolism, Transcription Factors metabolism, Breast Neoplasms genetics, Carcinogenesis genetics, DNA (Cytosine-5-)-Methyltransferases genetics, LIM-Homeodomain Proteins genetics, Mammary Glands, Animal metabolism, Mammary Neoplasms, Experimental genetics, Neoplastic Stem Cells metabolism, Transcription Factors genetics
- Abstract
Mammary stem/progenitor cells (MaSCs) maintain self-renewal of the mammary epithelium during puberty and pregnancy. DNA methylation provides a potential epigenetic mechanism for maintaining cellular memory during self-renewal. Although DNA methyltransferases (DNMTs) are dispensable for embryonic stem cell maintenance, their role in maintaining MaSCs and cancer stem cells (CSCs) in constantly replenishing mammary epithelium is unclear. Here we show that DNMT1 is indispensable for MaSC maintenance. Furthermore, we find that DNMT1 expression is elevated in mammary tumours, and mammary gland-specific DNMT1 deletion protects mice from mammary tumorigenesis by limiting the CSC pool. Through genome-scale methylation studies, we identify ISL1 as a direct DNMT1 target, hypermethylated and downregulated in mammary tumours and CSCs. DNMT inhibition or ISL1 expression in breast cancer cells limits CSC population. Altogether, our studies uncover an essential role for DNMT1 in MaSC and CSC maintenance and identify DNMT1-ISL1 axis as a potential therapeutic target for breast cancer treatment.
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- 2015
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13. Histone-fold domain protein NF-Y promotes chromatin accessibility for cell type-specific master transcription factors.
- Author
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Oldfield AJ, Yang P, Conway AE, Cinghu S, Freudenberg JM, Yellaboina S, and Jothi R
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- Animals, Binding Sites, CCAAT-Binding Factor metabolism, Cells, Cultured, Embryonic Stem Cells metabolism, Embryonic Stem Cells ultrastructure, Mice, Models, Genetic, Nucleosomes chemistry, Nucleosomes metabolism, Pluripotent Stem Cells, Transcription Factors chemistry, Transcription Factors metabolism, Transcription Factors physiology, CCAAT-Binding Factor physiology, Chromatin metabolism, Histones metabolism
- Abstract
Cell type-specific master transcription factors (TFs) play vital roles in defining cell identity and function. However, the roles ubiquitous factors play in the specification of cell identity remain underappreciated. Here we show that the ubiquitous CCAAT-binding NF-Y complex is required for the maintenance of embryonic stem cell (ESC) identity and is an essential component of the core pluripotency network. Genome-wide studies in ESCs and neurons reveal that NF-Y regulates not only genes with housekeeping functions through cell type-invariant promoter-proximal binding, but also genes required for cell identity by binding to cell type-specific enhancers with master TFs. Mechanistically, NF-Y's distinct DNA-binding mode promotes master/pioneer TF binding at enhancers by facilitating a permissive chromatin conformation. Our studies unearth a conceptually unique function for histone-fold domain (HFD) protein NF-Y in promoting chromatin accessibility and suggest that other HFD proteins with analogous structural and DNA-binding properties may function in similar ways., (Copyright © 2014 Elsevier Inc. All rights reserved.)
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- 2014
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14. Integrative framework for identification of key cell identity genes uncovers determinants of ES cell identity and homeostasis.
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Cinghu S, Yellaboina S, Freudenberg JM, Ghosh S, Zheng X, Oldfield AJ, Lackford BL, Zaykin DV, Hu G, and Jothi R
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- Animals, Cell Differentiation genetics, Cell Proliferation, Gene Expression Regulation, Homeodomain Proteins metabolism, Mice, Nanog Homeobox Protein, Oxidative Stress genetics, Phosphoproteins genetics, Phosphoproteins metabolism, Pluripotent Stem Cells cytology, Pluripotent Stem Cells metabolism, RNA Interference, RNA-Binding Proteins genetics, RNA-Binding Proteins metabolism, Reactive Oxygen Species metabolism, Reproducibility of Results, Transcription, Genetic, Tumor Suppressor Protein p53 metabolism, Nucleolin, Embryonic Stem Cells cytology, Embryonic Stem Cells metabolism, Homeostasis genetics
- Abstract
Identification of genes associated with specific biological phenotypes is a fundamental step toward understanding the molecular basis underlying development and pathogenesis. Although RNAi-based high-throughput screens are routinely used for this task, false discovery and sensitivity remain a challenge. Here we describe a computational framework for systematic integration of published gene expression data to identify genes defining a phenotype of interest. We applied our approach to rank-order all genes based on their likelihood of determining ES cell (ESC) identity. RNAi-mediated loss-of-function experiments on top-ranked genes unearthed many novel determinants of ESC identity, thus validating the derived gene ranks to serve as a rich and valuable resource for those working to uncover novel ESC regulators. Underscoring the value of our gene ranks, functional studies of our top-hit Nucleolin (Ncl), abundant in stem and cancer cells, revealed Ncl's essential role in the maintenance of ESC homeostasis by shielding against differentiation-inducing redox imbalance-induced oxidative stress. Notably, we report a conceptually novel mechanism involving a Nucleolin-dependent Nanog-p53 bistable switch regulating the homeostatic balance between self-renewal and differentiation in ESCs. Our findings connect the dots on a previously unknown regulatory circuitry involving genes associated with traits in both ESCs and cancer and might have profound implications for understanding cell fate decisions in cancer stem cells. The proposed computational framework, by helping to prioritize and preselect candidate genes for tests using complex and expensive genetic screens, provides a powerful yet inexpensive means for identification of key cell identity genes.
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- 2014
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15. MicroRNA-34c inversely couples the biological functions of the runt-related transcription factor RUNX2 and the tumor suppressor p53 in osteosarcoma.
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van der Deen M, Taipaleenmäki H, Zhang Y, Teplyuk NM, Gupta A, Cinghu S, Shogren K, Maran A, Yaszemski MJ, Ling L, Cool SM, Leong DT, Dierkes C, Zustin J, Salto-Tellez M, Ito Y, Bae SC, Zielenska M, Squire JA, Lian JB, Stein JL, Zambetti GP, Jones SN, Galindo M, Hesse E, Stein GS, and van Wijnen AJ
- Subjects
- Animals, Bone Neoplasms genetics, Bone Neoplasms pathology, Cell Cycle genetics, Cell Cycle radiation effects, Cell Line, Tumor, Cell Proliferation radiation effects, Core Binding Factor Alpha 1 Subunit genetics, Cyclin-Dependent Kinase Inhibitor p21 metabolism, DNA Damage, Down-Regulation genetics, Down-Regulation radiation effects, Gamma Rays, Gene Expression Regulation, Neoplastic radiation effects, Humans, Mice, Osteosarcoma genetics, Osteosarcoma pathology, Protein Stability radiation effects, RNA, Messenger genetics, RNA, Messenger metabolism, Tumor Suppressor Protein p14ARF metabolism, Tumor Suppressor Protein p53 deficiency, Bone Neoplasms metabolism, Core Binding Factor Alpha 1 Subunit metabolism, MicroRNAs metabolism, Osteosarcoma metabolism, Tumor Suppressor Protein p53 metabolism
- Abstract
Osteosarcoma (OS) is a primary bone tumor that is most prevalent during adolescence. RUNX2, which stimulates differentiation and suppresses proliferation of osteoblasts, is deregulated in OS. Here, we define pathological roles of RUNX2 in the etiology of OS and mechanisms by which RUNX2 expression is stimulated. RUNX2 is often highly expressed in human OS biopsies and cell lines. Small interference RNA-mediated depletion of RUNX2 inhibits growth of U2OS OS cells. RUNX2 levels are inversely linked to loss of p53 (which predisposes to OS) in distinct OS cell lines and osteoblasts. RUNX2 protein levels decrease upon stabilization of p53 with the MDM2 inhibitor Nutlin-3. Elevated RUNX2 protein expression is post-transcriptionally regulated and directly linked to diminished expression of several validated RUNX2 targeting microRNAs in human OS cells compared with mesenchymal progenitor cells. The p53-dependent miR-34c is the most significantly down-regulated RUNX2 targeting microRNAs in OS. Exogenous supplementation of miR-34c markedly decreases RUNX2 protein levels, whereas 3'-UTR reporter assays establish RUNX2 as a direct target of miR-34c in OS cells. Importantly, Nutlin-3-mediated stabilization of p53 increases expression of miR-34c and decreases RUNX2. Thus, a novel p53-miR-34c-RUNX2 network controls cell growth of osseous cells and is compromised in OS.
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- 2013
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16. Phosphorylation of the gastric tumor suppressor RUNX3 following H. pylori infection results in its localization to the cytoplasm.
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Cinghu S, Goh YM, Oh BC, Lee YS, Lee OJ, Devaraj H, and Bae SC
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- Animals, Cell Line, Tumor, Core Binding Factor Alpha 3 Subunit genetics, Cytoplasm microbiology, Cytoplasm pathology, Female, Gastric Mucosa metabolism, Gastric Mucosa microbiology, Gastric Mucosa pathology, Gastritis genetics, Gastritis pathology, Helicobacter Infections genetics, Helicobacter Infections pathology, Humans, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Phosphorylation genetics, Stomach Neoplasms microbiology, Stomach Neoplasms pathology, Stomach Neoplasms prevention & control, Core Binding Factor Alpha 3 Subunit metabolism, Cytoplasm metabolism, Gastritis metabolism, Helicobacter Infections metabolism, Helicobacter pylori
- Abstract
As H. pylori infection progresses, intestinal metaplasia (IM), a key event in gastric carcinogenesis, develops in the stomach. The mechanism by which H. pylori infection causes the trans-differentiation of gastric cells to intestinal-type cells remains an important question. In the current study, we found that RUNX3 is deregulated in all human IM specimens examined by either down regulation or mislocalization; Aberrant localization of a gastric tumor suppressor RUNX3 is observed in most human cases of IM with concurrent H. pylori infection, and RUNX3 is down-regulated in most cases of IM without H. pylori-infection. The cytoplasmic mislocalization of a RUNX3 was associated with H. pylori-induced c-Src activation and RUNX tyrosine phosphorylation. Moreover, gastric epithelial cells of Runx3(-/-) mice expressed the intestinal markers Muc2 and Li-Cadherin, which suggests that the deregulation of Runx3 is a key event in the intestinalization of the gastric epithelium. Collectively, the results of the current study suggest that RUNX3 deregulation is associated with H. pylori-induced pathogenesis and the development of IM., (Copyright © 2011 Wiley Periodicals, Inc.)
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- 2012
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17. Fused Toes Homolog modulates radiation cytotoxicity in uterine cervical cancer cells.
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Anandharaj A, Cinghu S, Kim WD, Yu JR, and Park WY
- Subjects
- Annexin A5 metabolism, Caspase 3 metabolism, Cell Cycle Proteins metabolism, Cell Death, Cell Line, Tumor, Cell Survival, Clone Cells, Down-Regulation, Female, Flow Cytometry, G1 Phase, Gene Knockdown Techniques, Humans, Immunohistochemistry, Poly(ADP-ribose) Polymerases metabolism, Protein Transport, Proto-Oncogene Proteins c-akt metabolism, Uterine Cervical Neoplasms enzymology, Adaptor Proteins, Signal Transducing metabolism, Apoptosis Regulatory Proteins metabolism, Radiation, Radiation Tolerance, Uterine Cervical Neoplasms metabolism, Uterine Cervical Neoplasms pathology
- Abstract
Radiotherapy is the major treatment modality for uterine cervical cancer, but in some cases, the disease is radioresistant. Defining the molecular events that contribute to radioresistance and progression of cancer are of critical importance. Here we evaluated the role of Fused Toes Homolog (FTS) in radiation resistance of cervical carcinoma. Immunostaning of cervical cancer cells and tissues revealed that FTS localization and expression was changed after radiation. Targeted stable knockdown of FTS in HeLa cells led to the growth inhibition after radiation. Radiation induced AKT mediated cytoprotective effect was countered by FTS knockdown which leads to PARP cleavage and caspase-3 activation leading to cell death. FTS knockdown promotes radiation induced cell cycle arrest at G0/G1 and apoptosis of HeLa cells with concurrent alterations in the display of cell cycle regulatory proteins. This study revealed FTS is involved in radioresistance of cervical cancer. Targeted inhibition of FTS led to the shutdown of key elemental characteristics of cervical cancer and could lead to an effective therapeutic strategy.
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- 2011
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18. FTS (fused toes homolog) a novel oncoprotein involved in uterine cervical carcinogenesis and a potential diagnostic marker for cervical cancer.
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Cinghu S, Anandharaj A, Lee HC, Yu JR, and Park WY
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- Adaptor Proteins, Signal Transducing antagonists & inhibitors, Apoptosis, Apoptosis Regulatory Proteins antagonists & inhibitors, Cell Line, Tumor, Cell Proliferation, Cell Survival, Cyclin-Dependent Kinase Inhibitor p21 genetics, Cyclin-Dependent Kinase Inhibitor p21 metabolism, Female, G1 Phase, Gene Knockdown Techniques, Humans, Precancerous Conditions metabolism, Resting Phase, Cell Cycle, Transcription, Genetic, Uterine Cervical Neoplasms metabolism, Adaptor Proteins, Signal Transducing metabolism, Apoptosis Regulatory Proteins metabolism, Biomarkers, Tumor metabolism, Precancerous Conditions diagnosis, Precancerous Conditions pathology, Uterine Cervical Neoplasms diagnosis, Uterine Cervical Neoplasms pathology
- Abstract
The high incidence and fatality rate of uterine cervical cancer warrant effective diagnostic and therapeutic target identification for this disease. Here, we have found a novel oncoprotein FTS (Fused Toes Homolog), which is involved in cervical cancer pathogenesis. Immunohistochemical analysis of human cervical biopsy samples revealed that the expression of FTS is absent in normal cervical epithelium but progressively overexpressed in human cervical intraneoplastic lesions (CIN-I to CIN-III), this characteristic phenomenon put this protein, a potential diagnostic marker for the screening of early neoplastic changes of cervix. Using FTS-specific small hairpin RNA (shRNA) in cervical cancer cells, we determined a specific role for FTS protein in, cervical neoplasia. Targeted stable knock down of FTS in HeLa cells led to the growth inhibition, cell-cycle arrest, and apoptosis with concurrent increase in p21 protein. FTS effectively represses the p21 mRNA expression in dual luciferase assay which indicates that p21 is transcriptionally regulated by this oncoprotein which in turn affect the regular cell-cycle process and its components. Consistent with this we found a reciprocal association between these proteins in early cervical neoplastic tissues. These data unraveled the involvement of new oncoprotein FTS in cervical cancer which plays a central role in carcinogenesis. Targeted inhibition of FTS lead to the shutdown of key elemental characteristics of cervical cancer and could lead to an effective therapeutic strategy for cervical cancer., (Copyright © 2010 Wiley-Liss, Inc.)
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- 2011
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19. Rapamycin-mediated mTOR inhibition attenuates survivin and sensitizes glioblastoma cells to radiation therapy.
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Anandharaj A, Cinghu S, and Park WY
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- Antibiotics, Antineoplastic pharmacology, Apoptosis drug effects, Apoptosis radiation effects, Blotting, Western, Cell Cycle drug effects, Cell Cycle radiation effects, Cell Line, Tumor, Cell Survival drug effects, Cell Survival radiation effects, Cyclin-Dependent Kinase Inhibitor p21 metabolism, Cyclin-Dependent Kinase Inhibitor p27 metabolism, Flow Cytometry, G1 Phase drug effects, G1 Phase radiation effects, Glioblastoma metabolism, Glioblastoma pathology, Histones metabolism, Humans, Phosphorylation drug effects, Phosphorylation radiation effects, Proto-Oncogene Proteins c-akt metabolism, Signal Transduction drug effects, Signal Transduction radiation effects, Survivin, TOR Serine-Threonine Kinases metabolism, Time Factors, Inhibitor of Apoptosis Proteins metabolism, Sirolimus pharmacology, TOR Serine-Threonine Kinases antagonists & inhibitors
- Abstract
Survivin, an antiapoptotic protein, is elevated in most malignancies and attributes to radiation resistance in tumors including glioblastoma multiforme. The downregulation of survivin could sensitize glioblastoma cells to radiation therapy. In this study, we investigated the effect of rapamycin, an inhibitor of mammalian target of rapamycin (mTOR), in attenuating survivin and enhancing the therapeutic efficacy for glioblastoma cells, and elucidated the underlying mechanisms. Here we tested various concentrations of rapamycin (1-8 nM) in combination with radiation dose 4 Gy. Rapamycin effectively modulated the protein kinase B (Akt)/mTOR pathway by inhibiting the phosphorylation of Akt and mTOR proteins, and this inhibition was further enhanced by radiation. The expression level of survivin was decreased in rapamycin pre-treatment glioblastoma cells followed by radiation; meanwhile, the phosphorylation of H2A histone family member X (H2AX) at serine-139 (γ-H2AX) was increased. p21 protein was also induced on radiation with rapamycin pre-treatment, which enhanced G1 arrest and the accumulation of cells at G0/subG1 phase. Furthermore, the clonogenic cell survival assay revealed a significant dose-dependent decrease in the surviving fraction for all three cell lines pre-treated with rapamycin. Our studies demonstrated that targeting survivin may be an effective approach for radiosensitization of malignant glioblastoma.
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- 2011
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20. Helicase-like transcription factor confers radiation resistance in cervical cancer through enhancing the DNA damage repair capacity.
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Cho S, Cinghu S, Yu JR, and Park WY
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- Apoptosis, Cell Proliferation, DNA Damage, DNA-Binding Proteins analysis, Female, HeLa Cells, Humans, RNA, Small Interfering genetics, Transcription Factors analysis, Uterine Cervical Neoplasms pathology, DNA Repair, DNA-Binding Proteins physiology, Radiation Tolerance, Transcription Factors physiology, Uterine Cervical Neoplasms radiotherapy
- Abstract
Helicase-like transcription factor (HLTF) is a member of the SWI/SNF (mating type switching/sucrose non-fermenting) family of ATPases/helicases and also has a RING-finger motif characteristic of ubiquitin ligase proteins. These features have led to suggestions that HLTF functions like yeast Rad5, which promotes replication through DNA lesions via a post-replication repair pathway. However, the function of HLTF in higher eukaryotes is still unknown. Herein, we found the overexpression of HLTF in radiation recurrent human uterine cervical carcinoma tissues when compared to disease free survived patients tissues. In this study, we used RNA interference techniques to investigate the potential function of HLTF in cervical cancer cell line HeLa and found that the cell proliferation was reduced by knockdown (KD) of HLTF. A host-cell reactivation assay showed that the capacity for repair to DNA damage induced by X-ray irradiation was reduced in HLTF KD cells. X-rays also increased apoptosis in HLTF KD cells. These results suggest that HLTF is involved in DNA repair and apoptosis in cancer cells, which might represent a target for gene therapies of human cancer.
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- 2011
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21. Src kinase phosphorylates RUNX3 at tyrosine residues and localizes the protein in the cytoplasm.
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Goh YM, Cinghu S, Hong ETH, Lee YS, Kim JH, Jang JW, Li YH, Chi XZ, Lee KS, Wee H, Ito Y, Oh BC, and Bae SC
- Subjects
- Breast Neoplasms metabolism, Cell Line, Tumor, Cell Nucleus metabolism, Cytoplasm metabolism, HeLa Cells, Humans, Phosphorylation, Protein Transport, RNA, Small Interfering metabolism, Stomach Neoplasms metabolism, Tyrosine genetics, Tyrosine metabolism, Core Binding Factor Alpha 3 Subunit metabolism, Gene Expression Regulation, Neoplastic, Tyrosine chemistry, src-Family Kinases metabolism
- Abstract
RUNX3 is a transcription factor that functions as a tumor suppressor. In some cancers, RUNX3 expression is down-regulated, usually due to promoter hypermethylation. Recently, it was found that RUNX3 can also be inactivated by the mislocalization of the protein in the cytoplasm. The molecular mechanisms controlling this mislocalization are poorly understood. In this study, we found that the overexpression of Src results in the tyrosine phosphorylation and cytoplasmic localization of RUNX3. We also found that the tyrosine residues of endogenous RUNX3 are phosphorylated and that the protein is localized in the cytoplasm in Src-activated cancer cell lines. We further showed that the knockdown of Src by small interfering RNA, or the inhibition of Src kinase activity by a chemical inhibitor, causes the re-localization of RUNX3 to the nucleus. Collectively, our results demonstrate that the tyrosine phosphorylation of RUNX3 by activated Src is associated with the cytoplasmic localization of RUNX3 in gastric and breast cancers.
- Published
- 2010
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22. Jab1/CSN5 induces the cytoplasmic localization and degradation of RUNX3.
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Kim JH, Choi JK, Cinghu S, Jang JW, Lee YS, Li YH, Goh YM, Chi XZ, Lee KS, Wee H, and Bae SC
- Subjects
- Active Transport, Cell Nucleus physiology, COP9 Signalosome Complex, Cell Nucleus metabolism, Cells, Cultured, HeLa Cells, Humans, Transcription, Genetic, Transfection, Core Binding Factor Alpha 3 Subunit analysis, Core Binding Factor Alpha 3 Subunit metabolism, Cytoplasm enzymology, Intracellular Signaling Peptides and Proteins metabolism, Peptide Hydrolases metabolism
- Abstract
Runt-related (RUNX) transcription factors play pivotal roles in neoplastic development and have tissue-specific developmental roles in hematopoiesis (RUNX1), osteogenesis (RUNX2), as well as neurogenesis and thymopoiesis (RUNX3). RUNX3 is a tumor suppressor in gastric carcinoma, and its expression is frequently inactivated by DNA methylation or its protein mislocalized in many cancer types, including gastric and breast cancer. Jun-activation domain-binding protein 1 (Jab1/CSN5), a component of the COP9 signalosome (CSN), is critical for nuclear export and the degradation of several tumor suppressor proteins, including p53, p27(Kip1), and Smad4. Here, we find that Jab1 facilitates nuclear export of RUNX3 that is controlled by CSN-associated kinases. RUNX3 sequestered in the cytoplasm is rapidly degraded through a proteasome-mediated pathway. Our results identify a novel mechanism of regulating nuclear export and protein stability of RUNX3 by the CSN complex., ((c) 2009 Wiley-Liss, Inc.)
- Published
- 2009
- Full Text
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