DOCSLIB.ORG

Transgelin 2 Promotes Paclitaxel Resistance, Migration, and Invasion

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Transgelin 2 Promotes Paclitaxel Resistance, Migration, and Invasion Published OnlineFirst September 5, 2019; DOI: 10.1158/1535-7163.MCT-19-0261 Cancer Biology and Translational Studies Molecular Cancer Therapeutics Transgelin 2 Promotes Paclitaxel Resistance, Migration, and Invasion of Breast Cancer by Directly Interacting with PTEN and Activating PI3K/Akt/GSK-3b Pathway Leichao Liu1, Ti Meng1, Xiaowei Zheng2, Yang Liu1, Ruifang Hao1, Yan Yan1,3, Siying Chen1, Haisheng You1, Jianfeng Xing3, and Yalin Dong1 Abstract MDR and tumor migration and invasion are still the main breast cancer cells were also enhanced by Transgelin 2 over- obstacles to effective breast cancer chemotherapies. Transgelin expression in vivo. Moreover, Transgelin 2 overexpression 2 has recently been shown to induce drug resistance, tumor activated the PI3K/Akt/GSK-3b pathway by increasing the migration, and invasion. The aim of this study was to deter- phosphorylation levels of Akt and GSK-3b and decreasing mine the biological functions of Transgelin 2 and the mech- the expression of PTEN. We also found that Transgelin 2 anism underlying how Transgelin 2 induces paclitaxel (PTX) could directly interact with PTEN and was located upstream resistance and the migration and invasion of breast cancer. We of PTEN. Furthermore, the PI3K/Akt pathway inhibitor detected that the protein level of Transgelin 2 was significantly MK-2206 reversed the resistance to paclitaxel and inhibited upregulated in breast cancer tissues compared with adjacent the migration and invasion of breast cancer cells. These nontumor tissues. A bioinformatics analysis indicated that findings indicate that Transgelin 2 promotes paclitaxel Transgelin 2 was significantly related to clinicopathologic resistance and the migration and invasion of breast cancer parameters and patient prognosis. Overexpression of Trans- by directly interacting with PTEN and activating the gelin 2 enhanced the migration and invasion of human breast PI3K/Akt/GSK-3b pathway. Transgelin 2 may therefore be cancer cells and decreased the sensitivity of breast cancer cells useful as a novel biomarker and therapeutic target for to paclitaxel. Meanwhile, the tumorigenesis and metastasis of breast cancer. Introduction molecular mechanisms of MDR and tumor metastasis is signif- icant for patients with breast cancer to develop new effective Breast cancer is the most common malignancy among women therapeutic strategies. worldwide (1). Although treatment with a combination of che- Paclitaxel (PTX) is widely applied in first-line chemotherapies motherapy and surgery has made remarkable progress in patients for treating breast cancer (4, 5). However, the efficacy of PTX is with breast cancer during the past few years, the recurrence and often hampered to acquired resistance in patients. Various cancer death remain very high due to high frequency of metastasis mechanisms have been proposed to explain PTX resistance, and recurrence. Even though additional drugs were used, breast including alterations in the level of expressed tubulin, overexpres- cancer cells generally gained MDR, finally causing tumor migra- sion of ATP-binding cassette (ABC) transporter proteins, and tion and invasion over time (2, 3). Therefore, exploring the apoptotic modulation (6–8). Several studies have found that the epithelial–mesenchymal transition (EMT) is related to acquisition of the MDR phenotype (9–11). The EMT is known to be involved in cancer 1 fi Department of Pharmacy, The First Af liated Hospital of Xi'an Jiaotong Uni- metastasis and is a process in which epithelial cancer cells lose versity, Xi';an, Shaanxi, P.R. China. 2Department of Pharmacy, The First Hospital of Xi'an, Xi'an, Shaanxi, P.R. China. 3School of pharmacy, Xi'an Jiaotong Uni- their typical epithelial characteristics and acquire mesenchymal versity, Xi'an, Shaanxi, P.R. China. traits. Cancer cells undergoing the EMT become migratory and invasive, enabling metastasis and chemotherapy resistance. Note: Supplementary data for this article are available at Molecular Cancer Therapeutics Online (http://mct.aacrjournals.org/). Therefore, identifying the molecular mechanism underlying tumor progression may ultimately lead to innovative therapeutic L. Liu and T. Meng contributed equally to this article. strategies against breast cancer. Corresponding Authors: Yalin Dong, The First Affiliated Hospital of Xi'an Transgelin 2, which is encoded by TAGLN2, belongs to the Jiaotong University, Xi'an, Shaanxi 710061, P.R. China. Phone: 8629-8532- family of actin-binding proteins and is a 22,391-Da protein 3241; Fax: 8629-8532-3240; E-mail: [email protected]; Jianfeng Xing, containing 199 amino acids. Deregulation of Transgelin 2 is School of Pharmacy, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China. Phone: 8629-8265-5139; E-mail: [email protected] considered to be associated with tumorigenesis and tumor devel- opment in various human malignancies (12–14). We previously Mol Cancer Ther 2019;18:2457–68 used proteomic technologies to reveal the expression of Transge- doi: 10.1158/1535-7163.MCT-19-0261 lin 2 to be 15.48-fold higher in PTX-resistant human breast cancer Ó2019 American Association for Cancer Research. cells (MCF-7/PTX) established by our laboratory than in breast www.aacrjournals.org 2457 Downloaded from mct.aacrjournals.org on September 30, 2021. © 2019 American Association for Cancer Research. Published OnlineFirst September 5, 2019; DOI: 10.1158/1535-7163.MCT-19-0261 Liu et al. cancer drug-sensitive cells (MCF-7/S; ref. 15). Furthermore, our Biochemistry and Cell Biology, Chinese Academy of Sciences, data demonstrated that Transgelin 2 can modulate the expression and MCF-7/PTX cell line was established as previously levels of MDR1, MRP1, and BCRP, suggesting that Transgelin 2 is described (19). All cell lines were grown in DMEM supple- critical for the occurrence of MDR in breast cancer cells (16). mented with 10% FBS, 1% penicillin, and streptomycin at 37C However, whether Transgelin 2 mediates the migration and in a humidified atmosphere of 5% CO2. The other culture invasion of breast cancer cells remains unclear. conditions of MCF-7/PTX cells werethesameastheMCF-7/S The PI3K/Akt/GSK-3b pathway is known to participate in the cells except for maintaining in 30 nmol/L paclitaxel (19). Cells regulation of various biological processes, such as the inhibition in exponential phase growth were observed under the inverted of apoptosis, induction of cell proliferation, and promotion of light microscope (Olympus). cancer metastasis (17). Our previous study demonstrated that Transgelin 2 activated the PI3K/Akt pathway in MCF-7/PTX cells, siRNA transfection, plasmid transient transfection, and thereby inhibiting mitochondrial apoptosis and inducing breast lentiviral infection cancer resistance (18). Nevertheless, the role of Transgelin 2 in the MCF-7/PTX cells were seeded in 6-well plate (5 Â 105 cells/ PI3K/Akt/GSK-3b pathway is still unknown and needs to be well). For siRNA transfection, cells were incubated in RPMI1640 explored further. without antibiotics for 24 hours, after which 0.1 nmol/L double- In this study, we found that Transgelin 2 was upregulated in stranded siRNA against TAGLN2 or nonspecific control siRNA breast cancer tissues, and that a strong expression of Transgelin 2 (GenePharma) was transfected with Lipofectamine 2000 (Invi- indicated a poor prognosis in patients with breast cancer. More- trogen). For plasmid transient transfection, 2 mg eukaryotic over, Transgelin 2 has a strong effect on the PTX resistance and the expression vector alone (pcDNA3.1) or pcDNA3.1-TAGLN2 invasion and metastasis of breast cancer cells both in vitro and (GenePharma) was transfected to MCF-7/S cells with the same in vivo. The molecular mechanism assays demonstrated that method. All the processes above were conducted according to the Transgelin 2 could suppress the expression of PTEN and activate manufacturer's instruction. the PI3K/Akt/GSK-3b pathway. MDA-MB-231 cells (2 Â 106 cells) were prepared and infected at a multiplicity of infection of 10 with negative control (Lenti-EV) or Transgelin 2-overexpressing lentiviruses (Lenti-TAGLN2) for Materials and Methods 10 hours at 37C in the presence of 5 mg/mL of polybrene. Chemicals and antibodies Selecting process was performed in 3 mg/mL puromycin 96 hours Paclitaxel and MK-2206 were purchased from Sike Pharma- after transfection. The lentivirus was purchased from GeneChem ceutical and Selleck Chemicals, respectively. 3-(4,5-Dimethylthia- and labeled with GFP. zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was from The expression level of Transgelin 2 and transfection efficiency Sigma. Annexin-V FLUOS staining kit was obtained from Invitro- were detected by qPCR and Western blot assays, supplemented by gen. The human monoclonal anti-N-cadherin, anti-P-gp, and fluorescent inverted microscope (ZEISS) to observe GFP expres- anti-BCRP were purchased from Abcam. The human monoclonal sion after lentiviral infection. anti-vimentin, anti-MRP1, and anti-Transgelin 2 antibodies were purchased from GeneTex. The human anti-Bcl-2 and anti-Bcl-2– MTT assay associated X (Bax) were from Epitomics. The human monoclonal Cells (5 Â 105/mL) were seeded in 96-well plates with different anti-E-cadherin, anti-PTEN, anti-Akt, anti-phospho-Akt (p-Akt), treatments for indicated duration. A solution of 20 mL of MTT anti-caspase-3, anti-caspase-9, anti-PARP, and anti-Snail were (5 mg/mL) was added into each well and incubated for 4 hours. purchased from Cell Signaling Technology. The human polyclon- Then 150 mL of dimethyl
Load more

Recommended publications
  • Transgelin Interacts with PARP1 and Affects Rho Signaling Pathway in Human Colon Cancer Cells
    Transgelin interacts with PARP1 and affects Rho signaling pathway in human colon cancer cells Zhen-xian Lew Guangzhou Concord Cancer Center Hui-min Zhou First Aliated Hospital of Guangdong Pharmaceutical College Yuan-yuan Fang Tongling Peoples's Hospital Zhen Ye Sun Yat-sen Memorial Hospital Wa Zhong Sun Yat-sen Memorial Hospital Xin-yi Yang The Seventh Aliated Hospital Sun Yat-sen University Zhong Yu Sun Yat-sen Memorial Hospital of Sun Yat-sen University Dan-yu Chen Sun Yat-sen Memorial Hospital Si-min Luo Sun Yat-sen Memorial Hospital Li-fei Chen Sun Yat-sen Memorial Hospital Ying Lin ( [email protected] ) Sun Yat-sen Memorial Hospital https://orcid.org/0000-0003-2416-2154 Primary research Keywords: Transgelin, PARP1, Colon Cancer, Rho Signaling, Bioinformatics Posted Date: July 1st, 2020 DOI: https://doi.org/10.21203/rs.3.rs-16964/v2 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/22 Version of Record: A version of this preprint was published on August 3rd, 2020. See the published version at https://doi.org/10.1186/s12935-020-01461-y. Page 2/22 Abstract Background: Transgelin, an actin-binding protein, is associated with the cytoskeleton remodeling. Our previous studies found that transgelin was up-regulated in node-positive colorectal cancer versus in node- negative disease. Over-expression of TAGLN affected the expression of 256 downstream transcripts and increased the metastatic potential of colon cancer cells in vitro and in vivo. This study aims to explore the mechanisms that transgelin participates in the metastasis of colon cancer cells.
    [Show full text]
  • Differential Expression of Drosophila Transgelins Throughout Development
    fcell-09-648568 July 6, 2021 Time: 18:30 # 1 ORIGINAL RESEARCH published: 12 July 2021 doi: 10.3389/fcell.2021.648568 Differential Expression of Drosophila Transgelins Throughout Development Katerina M. Vakaloglou1†, Maria Mouratidou1†, Athina Keramidioti1,2 and Christos G. Zervas1* 1 Center of Basic Research, Biomedical Research Foundation, Academy of Athens, Athens, Greece, 2 Department of Biochemistry and Biotechnology, University of Thessaly, Larissa, Greece Transgelins are a conserved family of actin-binding proteins involved in cytoskeletal remodeling, cell contractility, and cell shape. In both mammals and Drosophila, three genes encode transgelin proteins. Transgelins exhibit a broad and overlapping expression pattern, which has obscured the precise identification of their role in development. Here, we report the first systematic developmental analysis of all Drosophila transgelin proteins, namely, Mp20, CG5023, and Chd64 in the Edited by: living organism. Drosophila transgelins display overall higher sequence identity with Lei-Miao Yin, mammalian TAGLN-3 and TAGLN-2 than with TAGLN. Detailed examination in different Shanghai University of Traditional Chinese Medicine, China developmental stages revealed that Mp20 and CG5023 are predominantly expressed in Reviewed by: mesodermal tissues with the onset of myogenesis and accumulate in the cytoplasm Cedric Soler, of all somatic muscles and heart in the late embryo. Notably, at postembryonic Clermont Université, France Simone Diestel, developmental stages, Mp20 and CG5023 are detected in the gut’s circumferential University of Bonn, Germany muscles with distinct subcellular localization: Z-lines for Mp20 and sarcomere and Rajesh Gunage, nucleus for CG5023. Only CG5023 is strongly detected in the adult fly in the abdominal, Boston Children’s Hospital, United States leg, and synchronous thoracic muscles.
    [Show full text]
  • KIF23 Enhances Cell Proliferation in Pancreatic Ductal Adenocarcinoma and Is a Potent Therapeutic Target
    1394 Original Article Page 1 of 15 KIF23 enhances cell proliferation in pancreatic ductal adenocarcinoma and is a potent therapeutic target Chun-Tao Gao1#, Jin Ren2#, Jie Yu1,3#, Sheng-Nan Li1, Xiao-Fan Guo1, Yi-Zhang Zhou1 1Department of Pancreatic Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin Key Laboratory of Cancer Prevention and Therapy, Tianjin’s Clinical Research Center for Cancer, Tianjin, China; 2Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Taiyuan, China; 3The First Hospital of Shanxi Medical University, Taiyuan, China Contributions: (I) Conception and design: CT Gao, J Ren; (II) Administrative support: CT Gao; (III) Provision of study materials or patients: J Yu; (IV) Collection and assembly of data: J Ren, SN Li, YZ Zhou; (V) Data analysis and interpretation: XF Guo, J Ren; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors. #These authors contributed equally to this work. Correspondence to: Chun-Tao Gao. Department of Pancreatic Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin Key Laboratory of Cancer Prevention and Therapy, Tianjin’s Clinical Research Center for Cancer, Huan-hu-xi Road, He-xi District, Tianjin 300060, China. Email: [email protected]. Background: In recent research, high expression of kinesin family member 23 (KIF23), one of the kinesin motor proteins involved in the regulation of cytokinesis, has been shown to be related to poor prognosis in glioma and paclitaxel-resistant gastric cancer, as a results of the enhancement of proliferation, migration, and invasion. In this study, we analyzed the role of KIF23 in the progression of pancreatic ductal adenocarcinoma.
    [Show full text]
  • Polyclonal Antibody to Transgelin (TAGLN) (C-Term) - Aff - Purified
    OriGene Technologies, Inc. OriGene Technologies GmbH 9620 Medical Center Drive, Ste 200 Schillerstr. 5 Rockville, MD 20850 32052 Herford UNITED STATES GERMANY Phone: +1-888-267-4436 Phone: +49-5221-34606-0 Fax: +1-301-340-8606 Fax: +49-5221-34606-11 [email protected] [email protected] AP16212PU-N Polyclonal Antibody to Transgelin (TAGLN) (C-term) - Aff - Purified Alternate names: 22 kDa actin-binding protein, SM22, Smooth muscle protein 22-alpha, WS3-10 Quantity: 0.1 mg Concentration: 0.5 mg/ml Background: TAGLN is a transformation and shape-change sensitive actin cross-linking/gelling protein found in fibroblasts and smooth muscle. Its expression is down-regulated in many cell lines, and this down-regulation may be an early and sensitive marker for the onset of transformation. A functional role of this protein is unclear. Uniprot ID: Q01995 NCBI: NP_003177.2 GeneID: 6876 Host: Goat Immunogen: Peptide with sequence from the C-Terminus of the protein sequence according to NP_003177.2. Genename: TAGLN AA Sequence: C-MTGYGRPRQIIS Remarks: NP_003177.2 and NP_001001522.1 represent identical protein. Format: State: Liquid purified Ig fraction Purification: Ammonium Sulphate Precipitation followed by Antigen Affinity Chromatography using the immunizing peptide Buffer System: Tris saline, pH~7.3 Preservatives: 0.02% Sodium Azide Stabilizers: 0.5% BSA Applications: Peptide ELISA: 1/8000 (Detection Limit). Western blot: 0.01-0.03 mg/ml. Approx 24kDa band observed in Human Placenta lysates (calculated MW of 22.6kDa according to NP_003177.2). Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
    [Show full text]
  • Transcriptomic Landscape of Early Age Onset of Colorectal Cancer Identifies
    www.nature.com/scientificreports OPEN Transcriptomic landscape of early age onset of colorectal cancer identifes novel genes and pathways in Indian CRC patients Manish Pratap Singh, Sandhya Rai, Nand K. Singh & Sameer Srivastava* Past decades of the current millennium have witnessed an unprecedented rise in Early age Onset of Colo Rectal Cancer (EOCRC) cases in India as well as across the globe. Unfortunately, EOCRCs are diagnosed at a more advanced stage of cancer. Moreover, the aetiology of EOCRC is not fully explored and still remains obscure. This study is aimed towards the identifcation of genes and pathways implicated in the EOCRC. In the present study, we performed high throughput RNA sequencing of colorectal tumor tissues for four EOCRC (median age 43.5 years) samples with adjacent mucosa and performed subsequent bioinformatics analysis to identify novel deregulated pathways and genes. Our integrated analysis identifes 17 hub genes (INSR, TNS1, IL1RAP, CD22, FCRLA, CXCL3, HGF, MS4A1, CD79B, CXCR2, IL1A, PTPN11, IRS1, IL1B, MET, TCL1A, and IL1R1). Pathway analysis of identifed genes revealed that they were involved in the MAPK signaling pathway, hematopoietic cell lineage, cytokine–cytokine receptor pathway and PI3K-Akt signaling pathway. Survival and stage plot analysis identifed four genes CXCL3, IL1B, MET and TNS1 genes (p = 0.015, 0.038, 0.049 and 0.011 respectively), signifcantly associated with overall survival. Further, diferential expression of TNS1 and MET were confrmed on the validation cohort of the 5 EOCRCs (median age < 50 years and sporadic origin). This is the frst approach to fnd early age onset biomarkers in Indian CRC patients.
    [Show full text]
  • Monoclonal Antibody to Transgelin (SM22-Alpha)(Clone : TAGLN/247)
    9853 Pacific Heights Blvd. Suite D. San Diego, CA 92121, USA Tel: 858-263-4982 Email: [email protected] 36-1708: Monoclonal Antibody to Transgelin (SM22-alpha)(Clone : TAGLN/247) Clonality : Monoclonal Clone Name : TAGLN/247 Application : FACS,IF,IHC Reactivity : Human, Mouse Gene : TAGLN Gene ID : 6876 Uniprot ID : Q01995 Format : Purified Alternative Name : TAGLN,SM22,WS3-10 Isotype : Mouse IgG1 Kappa Immunogen Information : Recombinant full-length human transgelin (TAGLN) protein. Description This MAb recognizes a 22kDa protein, identified as Transgelin, also designated SM22-alpha. It may cross-react with SM22- beta. Transgelin is expressed abundantly in smooth muscle cells. The human transgelin gene encodes a 201 amino acid protein that contains nuclear factor-binding motifs known to regulate transcription in smooth muscle. During embryogenesis, transgelin is expressed in smooth, cardiac and skeletal muscle, but is restricted during late fetal development and adulthood to all vascular and visceral smooth muscle cells and low levels of expression in heart. Transgelin is down regulated in several transformed cell lines, indicating that a reduction of transgelin expression may be an early indicator of the onset of transformation. Transgelin also binds Actin, causing Actin fibers to gel within minutes of binding. Binding of transgelin to Actin occurs at a ratio of 1:6 Actin monomers. Product Info Amount : 100 µg Purification : Affinity Chromatography 100 µg in 500 µl PBS containing 0.05% BSA and 0.05% sodium azide. Sodium azide is highly Content : toxic. Store the antibody at 4°C; stable for 6 months. For long-term storage; store at -20°C. Avoid Storage condition : repeated freeze and thaw cycles.
    [Show full text]
  • Transgelin Increases Metastatic Potential of Colorectal Cancer Cells
    Zhou et al. BMC Cancer (2016) 16:55 DOI 10.1186/s12885-016-2105-8 RESEARCH ARTICLE Open Access Transgelin increases metastatic potential of colorectal cancer cells in vivo and alters expression of genes involved in cell motility Hui-min Zhou1,2,3, Yuan-yuan Fang1,2, Paul M. Weinberger4,5, Ling-ling Ding6, John K. Cowell5, Farlyn Z. Hudson7, Mingqiang Ren5, Jeffrey R. Lee8, Qi-kui Chen2, Hong Su2, William S. Dynan7,9* and Ying Lin1,2* Abstract Background: Transgelin is an actin-binding protein that promotes motility in normal cells. Although the role of transgelin in cancer is controversial, a number of studies have shown that elevated levels correlate with aggressive tumor behavior, advanced stage, and poor prognosis. Here we sought to determine the role of transgelin more directly by determining whether experimental manipulation of transgelin levels in colorectal cancer (CRC) cells led to changes in metastatic potential in vivo. Methods: Isogenic CRC cell lines that differ in transgelin expression were characterized using in vitro assays of growth and invasiveness and a mouse tail vein assay of experimental metastasis. Downstream effects of transgelin overexpression were investigated by gene expression profiling and quantitative PCR. Results: Stable overexpression of transgelin in RKO cells, which have low endogenous levels, led to increased invasiveness, growth at low density, and growth in soft agar. Overexpression also led to an increase in the number and size of lung metastases in the mouse tail vein injection model. Similarly, attenuation of transgelin expression in HCT116 cells, which have high endogenous levels, decreased metastases in the same model.
    [Show full text]
  • Transgelin Interacts with PARP1 in Human Colon Cancer Cells
    Lew et al. Cancer Cell Int (2020) 20:366 https://doi.org/10.1186/s12935-020-01461-y Cancer Cell International PRIMARY RESEARCH Open Access Transgelin interacts with PARP1 in human colon cancer cells Zhen‑xian Lew1,2,3†, Hui‑min Zhou4†, Yuan‑yuan Fang5, Zhen Ye1,2, Wa Zhong1,2, Xin‑yi Yang6, Zhong Yu1,2, Dan‑yu Chen1,2, Si‑min Luo1,2, Li‑fei Chen7 and Ying Lin1,2* Abstract Background: Transgelin, an actin‑binding protein, is associated with cytoskeleton remodeling. Findings from our previous studies demonstrated that transgelin was up‑regulated in node‑positive colorectal cancer (CRC) ver‑ sus node‑negative disease. Over‑expression of TAGLN afected the expression of 256 downstream transcripts and increased the metastatic potential of colon cancer cells in vitro and in vivo. This study aims to explore the mechanisms through which transgelin participates in the metastasis of colon cancer cells. Methods: Immunofuorescence and immunoblotting analysis were used to determine the cellular localization of endogenous and exogenous transgelin in colon cancer cells. Co‑immunoprecipitation and subsequently high‑ performance liquid chromatography/tandem mass spectrometry were performed to identify the proteins that were potentially interacting with transgelin. The 256 downstream transcripts regulated by transgelin were analyzed with bioinformatics methods to discriminate the specifc key genes and signaling pathways. The Gene‑Cloud of Biotech‑ nology Information (GCBI) tools were used to predict the potential transcription factors (TFs) for the key genes. The predicted TFs corresponded to the proteins identifed to interact with transgelin. The interaction between transgelin and the TFs was verifed by co‑immunoprecipitation and immunofuorescence.
    [Show full text]
  • Transgelin Is a Novel Marker of Smooth Muscle Differentiation That Improves Diagnostic Accuracy of Leiomyosarcomas
    Modern Pathology (2013) 26, 502–510 502 & 2013 USCAP, Inc All rights reserved 0893-3952/13 $32.00 Transgelin is a novel marker of smooth muscle differentiation that improves diagnostic accuracy of leiomyosarcomas: a comparative immunohistochemical reappraisal of myogenic markers in 900 soft tissue tumors Yves-Marie Robin1, Nicolas Penel2,3, Gae¨lle Pe´rot4,5, Agnes Neuville4,5,6, Vale´rie Ve´lasco4,5, Dominique Ranche`re-Vince7, Philippe Terrier8 and Jean-Michel Coindre4,5,6 1Department of Biology, Unit of Morphological and Molecular Pathology, Centre Oscar Lambret, Lille Cedex, France; 2Department of General Oncology, Centre Oscar Lambret, Lille Cedex, France; 3Research Unit (EA 2694), Medical School University, Lille-Nord-de-France University, Lille Cedex, France; 4Department of Pathology, Institut Bergonie´, Bordeaux Cedex, France; 5INSERM U916, Institut Bergonie´, Bordeaux, France; 6Laboratory of Pathology, Universite´ Victor Segalen Bordeaux 2, Bordeaux, France; 7Department of Patholoy, Centre Le´on Be´rard, Lyon, France and 8Department of Pathology, Institut Gustave Roussy, Villejuif, France Immunohistochemical use of myogenic markers serves to define smooth or skeletal muscle differentiation in soft tissue tumors. Establishing smooth muscle differentiation in malignant lesions can be challenging in some cases. We immunohistochemically examined 900 soft tissue tumors selected from the French Sarcoma Group’s archived tissue collection, which contains a large number of leiomyosarcomas. The four most widely used smooth muscle diagnostic markers were evaluated (smooth muscle actin, desmin, h-caldesmon and calponin), and compared with a novel marker, transgelin. The diagnostic performance of each marker was statistically assessed in terms of sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV) and accuracy (A), in leiomyosarcomas versus all other sarcomas including gastrointestinal stromal tumors (GIST), and second in leiomyosarcomas versus specific tumor types.
    [Show full text]
  • Characterization of Poldip2 Knockout Mice: Avoiding Incorrect Gene Targeting
    bioRxiv preprint doi: https://doi.org/10.1101/2021.02.02.429447; this version posted February 3, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Characterization of Poldip2 knockout mice: avoiding incorrect gene targeting Bernard Lassègue1*, Sandeep Kumar2, Rohan Mandavilli1, Keke Wang1, Michelle Tsai1, Dong-Won Kang2, Marina S. Hernandes1, Alejandra San Martín1, Hanjoong Jo1,2, W. Robert Taylor1,2,3 and Kathy K. Griendling1 1Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA 2Wallace H. Coulter Department of Biomedical Engineering, Emory University and Georgia Institute of Technology, Atlanta, GA 3Division of Cardiology, Atlanta VA Medical Center, Decatur, GA Running title: Poldip2 knockout mice Keywords: Poldip2, mouse, conditional knockout, constitutive knockout, gene targeting, ectopic targeting, gene duplication, unexpected mutation *Corresponding author: Bernard Lassègue Division of Cardiology Emory University School of Medicine 101 Woodruff Circle WMB 308B Atlanta, GA 30322 [email protected] bioRxiv preprint doi: https://doi.org/10.1101/2021.02.02.429447; this version posted February 3, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract POLDIP2 is a multifunctional protein whose roles are only partially understood. Our laboratory previously reported physiological studies performed using a mouse gene trap model, which suffered from two limitations: perinatal lethality in homozygotes and constitutive Poldip2 inactivation. To overcome these limitations, we developed a new conditional floxed Poldip2 model. The first part of the present study shows that our initial floxed mice were affected by an unexpected mutation, which was not readily detected by Southern blotting and traditional PCR.
    [Show full text]
  • Human Protein Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for AR09539PU-N Transgelin (TAGLN) (1-201, His-tag) Human Protein Product data: Product Type: Recombinant Proteins Description: Transgelin (TAGLN) (1-201, His-tag) human recombinant protein, 0.1 mg Species: Human Expression Host: E. coli Tag: His-tag Predicted MW: 24.8 kDa Concentration: lot specific Purity: >85% by SDS – PAGE Buffer: Presentation State: Purified State: Liquid purified protein Buffer System: 20mM Tris-HCl buffer (pH 8.0) containing 20% glycerol, 1mM DTT Preparation: Liquid purified protein Protein Description: Recombinant human TAGLN protein, fused to his-tag at N-terminus was expressed in E.coli and purified by using conventional chromatography techniques. Storage: Store undiluted at 2-8°C for up to two weeks or (in aliquots) at -20°C or -70°C for longer. Avoid repeated freezing and thawing. Stability: Shelf life: one year from despatch. RefSeq: NP_001001522 Locus ID: 6876 UniProt ID: Q01995, Q5U0D2 Cytogenetics: 11q23.3 Synonyms: SM22; SM22-alpha; SMCC; TAGLN1; WS3-10 This product is to be used for laboratory only. Not for diagnostic or therapeutic use. View online » ©2021 OriGene Technologies, Inc., 9620 Medical Center Drive, Ste 200, Rockville, MD 20850, US 1 / 2 Transgelin (TAGLN) (1-201, His-tag) Human Protein – AR09539PU-N Summary: This gene encodes a shape change and transformation sensitive actin-binding protein which belongs to the calponin family. It is ubiquitously expressed in vascular and visceral smooth muscle, and is an early marker of smooth muscle differentiation.
    [Show full text]
  • Identi Cation of Novel Hub Genes Through Expression Pro Les
    Identication of Novel Hub Genes Through Expression Proles Analysis in Adrenocortical Carcinoma Chenhe Yao Beijing Zhicheng Biomedical Technology Co,Ltd Xiaoling Zhao Beijing Zhicheng Biomedical Technology Co,Ltd Xuemeng Shang Beijing Zhicheng Biomedical Technology Co,Ltd Binghan Jia Beijing Zhicheng Biomedical Technology Co,Ltd Shuaijie Dou Beijing Zhicheng Biomedical Technology Co,Ltd Weiliang Ye Beijing Zhicheng Biomedical Technology Co,Ltd Yuemei Yang ( [email protected] ) Beijing Zhicheng Biomedical Technology,Co.,Ltd. Research Keywords: Adrenocortical carcinoma, biomarkers, gene expression proling, Co-expression, prognosis Posted Date: July 1st, 2020 DOI: https://doi.org/10.21203/rs.3.rs-38768/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/21 Abstract Background: Adrenocortical carcinoma (ACC) is a heterogeneous and rare malignant tumor associated with a poor prognosis. The molecular mechanisms of ACC remain elusive and more accurate biomarkers for the prediction of prognosis are needed. Methods: In this study, integrative proling analyses were performed to identify novel hub genes in ACC to provide promising targets for future investigation. Three gene expression proling datasets in the GEO database were used for the identication of overlapped differentially expressed genes (DEGs) following the criteria of adj.P.Value<0.05 and |log2 FC|>0.5 in ACC. Novel hub genes were screened out following a series of processes: the retrieval of DEGs with no known associations with ACC on Pubmed, then the cross-validation of expression values and signicant associations with overall survival in the GEPIA2 and starBase databases, and nally the prediction of gene-tumor association in the GeneCards database.
    [Show full text]