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사람의 침윤성 암세포에서 후성학적 BTG2/TIS21/PC3 발현 조절과 암 억제 기전 연구
No full textB cell translocation gene 2 (BTG2/TIS21/PC3) belongs to the family of antiproliferative (APRO) genes and reported as a tumor suppressor by our group and others. Expression of BTG2 is significantly reduced in cancers developed in various organs and tissues. EJ (bladder carcinoma cells), MKN-1 (gastric cancer cells) are highly invasive and metastatic cells with very less endogenous BTG2 expression due to epigenetic regulation. Significantly lower endogenous expression of BTG2 was observed in human muscle-invasive bladder cancers (MIBC) than matched normal tissues and non-muscle invasive bladder cancers (NMIBC). BTG2 expression was inversely correlated with increased expression of the DNA methyltransferases DNMT1 and DNMT3a in MIBC, but not NMIBC, suggesting a potential role for BTG2 expression in muscle invasion of bladder cancer. Over 90% of tumor tissues revealed strong methylation at CpG islands of the BTG2 gene, compared with no methylation in the normal tissues, implying epigenetic regulation of BTG2 expression in bladder carcinogenesis. BTG2 is constitutively expressed in mucous epithelium and parietal cells of gastric glands in stomach, and the expression was increased in mucous epithelium with H. pylori infection as opposed to loss in human gastric adenocarcinoma. Indeed, adenoviral transduction of BTG2 significantly inhibited Tipα activity in MKN-1 and MGT-40, human and mouse gastric cancer cells, respectively, thereby downregulated TNFα expression and Erk1/2 phosphorylation via reducing nucleolin, Tipα receptor, expression. Chromatin immunoprecipitation proved that BTG2 inhibited Sp1 expression and it’s binding to the promoter of nucleolin gene. In addition, BTG2 expression significantly reduced membrane localized nucleolin expression in cancer cells and the loss of BTG2/TIS21 expression rather induced cytoplasmic nucleolin availability in gastric cancer tissues, evidenced by immunoblot and immunohistochemistry. The higher expression of BTG2 and the lower nucleolin expression accompanied with the better overall survival of the poorly differentiated gastric cancer patients.B cell translocation gene 2(BTG2/TIS21/PC3)는 antiproliferative gene(APRO) 계열에 속하며 본 연구팀을 포함한 많은 연구자들에 의해 BTG2 의 암억제 기능이 연구되어 왔다. BTG2 는 여러 조직과 기관에서 발생한 암에서 그 발현이 감소되어 있고, 특히 침윤(invasion)과 전이(metastasis)를 잘하는 암세포인 EJ (bladder carcinoma cells)와 MKN-1 (gastric cancer cells)에서도 후성유전학적(epigenetic regulation)으로 BTG2 발현이 매우 감소되어 있다. 최근 사람에서, 이런 침윤성과 전이성이 높은 암에서 BTG2 가 침윤을 억제하고 침윤을 조장하는 단백질과 mRNA 발현과는 음의 상관도를 보인다는 보고가 많기에, 본 저자는 BTG2 가 어떤 기전으로 침윤성 암을 조절하는지를 중점적으로 연구하였다. 사람의 근육침범 방광암(muscleinvasive bladder cancers, MIBC)에서 비-근육침범 방광암(non-muscle invasive bladder cancers, NMIBC)보다 BTG2 의 발현이 현저히 감소되어 있다는 것을 발견하였다. 이런 MIBC 에서의 BTG2 의 감소는 반대로 DNA 메틸기전달효소 DNMT1 과 DNMT3a 의 증가와 연관이 있었고, 이런 음의 상관관계는 NMIBC 에서는 발견되지 않음으로써 BTG2 이 방광암의 근육 침범에 분명한 역할을 하고 있다는 가능성을 보였다. 방광암 조직들의 90%에서 BTG2 유전자의 CpG 섬에 강한 메틸화를 보였고, 반대로 정상 방광조직에서는 메틸화가 발견되지 않음으로써, 방광암발생과정에서 BTG2 이 후성학적 조절을 받는다는 사실도 증명했다. 한편, BTG2 는 위 점막의 상피세포와 위샘의 벽세포(Parietal cell)에서 기본적으로 발현되고 있으며, 이 유전자의 발현은 H. pylori 감염시 위 점막 상피세포에서 증가되고, 반대로 위선암(gastric adenocarcinoma)에서는 감소됨을 발견했다. 사람 암세포 MKN-1 과 쥐 암세포 MGT-40 에서 BTG2 를 adenoviral transduction 을 통해 발현시키면 Tumor necrosis factor- α inducing protein (Tip α ) 활성이 억제되었고, 이는 Tip α receptor 인 nucleolin 을 감소시킴으로 TNFα의 발현과 Erk1/2 의 인산화를 억제시켰다. BTG2 는 Sp1 의 발현과 또한 염색질 면역침전(Chromatin immunoprecipitation) 실험을 통해 nucleolin 의 promoter 에 결합하는 Sp1 도 감소시킴을 확인했다. 또한 암세포에서 BTG2 를 발현시키면 세포막에 위치한 nucleolin 의 발현이 감소되었고, 반대로 BTG2 발현을 감소시키면 세포질 내에 nucleolin 의 발현이 증가함을 면역조직화학법과 면역블롯검사를 통해 위암조직에서 밝혔다. 이런 BTG2 의 강한 발현과 nuclolin 의 발현 저하는 저분화 위암 환자에서의 높은 생존율(overall survival)과 연관이 있었다. 더욱이 이런 BTG2 의 발현증가는 침윤성 암세포에서도 암 진행과 성장을 억제했다. 따라서, 본 저자는 본 연구를 통해 BTG2 유전자가 침윤성 암 환자를 위한 강력한 치료로 사용될 수 있으리라 제안한다.ABSTRACT i
TABLE OF CONTENTS iii
LIST OF FIGURES vi
ABBREVIATIONS viii
I. INTRODUCTION 1
II. EXPERIMENTAL METHODS 7
A. Tissues and cell cultures 7
B. RNA isolation and reverse transcription 8
C. Real-time and RT-PCR analyses 8
D. Methylation-specific PCR (MSP) and unmethylation-specific PCR (USP) analyses 9
E. Western blotting 9
F. DNMT activity assay 10
G. Chromatin immunoprecipitation (ChIP) analysis 10
H. Cloning of CpG islands and sequencing analyses 10
I. Immunoprecipitation/immunoblot analysis 11
J. Regulation of gene expression 11
K. Tumorigenesis study 12
L. Invasion assay 13
M. Immunohistochemistry analysis 14
N. Preparation of recombinant Tipα protein 15
O. Subcellular fractionation 15
P. Statistical analysis 15
III. RESULTS 16
A. Downregulation of BTG2 expression in human MIBC by DNA methylation of BTG2 gene 16
B. Inverse correlations between the expressions of BTG2 vs DNMT1 and DNMT3a in MIBC 18
C. Upregulation of BTG2 expression in EJ bladder cancer cells upon Decitabine treatment 22
D. Chromatin remodeling at the promoter and intron of BTG2 gene after decitabine treatment 27
E. Sp1, the transcription factor, for BTG2 gene upon decitabine treatment 28
F. Induction of BTG2 expression by knockdown of DNMT1 in EJ cells 32
G. Downregulation of tumorigenesis and cell cycle arrest by BTG2 overexpression 35
H. Inhibition of tumor invasion by BTG2 via downregulation of DNMT1
expression 38
I. Direct effect of BTG2/TIS21 expression on the inhibition of cancer invasiveness 41
J. Immunohistochemical findings 44
K. Induction of BTG2 expression by decitabine treatment in various cancer cell lines in addition to APRO gene expression analyses 46
L. Expression of BTG2/TIS21 is increased in mucous epithelium infected with H. pylori, but lost in human gastric adenocarcinoma 48
M. Absence of endogenous BTG2/TIS21 expression in the gastric cancer cells 51
N. Expression of BTG2/TIS21 is epigenetically regulated in gastric adenocarcinoma 52
O. Inhibition of cancer cell proliferation by BTG2/TIS21 gene 54
P. Inhibition of Tipα activity by BTG2/TIS21 in human gastric cancer cells 55
Q. Inhibition of Tipα activity by downregulation p-ERK1/2 in human gastric cancer cells 57
R. Downregulation of nucleolin expression by BTG2/TIS21 59
S. Expression of nucleolin, Tipα receptor, was reduced by BTG2 expression via inhibiting Sp1binding to the nucleolin promoter 61
T. Inverse regulation of Tipα -induced TNFα expression by NCL and BTG2 64
U. Reciprocal expression of BTG2/TIS21 and nucleolin expressions in normal and cancer regions 68
V. Inverse regulation of overall survival of gastric cancer patients by BTG2/TIS21 and NCL genes 71
IV. DISCUSSION 74
V. SUMMARY 80
VI. CONCLUSIONS 82
VII. REFERENCES 83
국문요약 95Docto
Dr. Duane M. Jackson, Morehouse College, July 2011
No full textThis video is a conversation with Dr. Duane M. Jackson. Dr. Jackson talks about his paper, "Recall and the Serial Position Effect: The Role of Primacy and Recency on Accounting Students' Performance." Jackie Daniel, AUC Woodruff Library, is the interviewer
"Reflections on the subject of Emigration from Europe with a view to Settlement in the United States" By M. Carey.
No full text"Reflections on the subject of Emigration from Europe with a view to Settlement in the United States: containing bried sketches of the moral and political character of those states.
By M. Carey, member of the American philosophical, and of the American Antiquarian Society, and author of The Olive Branch, Cindiciae Hibernicae, essays on banking, on political economy, and on internal improvement.
To which are now added the English editor's comments on the subject; together with Important Advice to Emigrants, and Cautions Against Impositions Practiced in the Outports
Dispelling the Myths Behind First-author Citation Counts
No full textWe conducted a full-scale evaluative citation analysis study of scholars in the XML research field to explore just how different from each other author rankings resulting from different citation counting methods actually are, and to demonstrate the capability of emerging data and tools on the Web in supporting more realistic citation counting methods. Our results contest some common arguments for the continued
use of first-author citation counts in the evaluation of scholars, such as high correlations between author rankings by first-author citation counts and other citation
counting methods, and high costs of using more realistic citation counting methods that are not well-supported by the ISI databases. It is argued that increasingly available digital full text research papers make it possible for citation analysis studies to go beyond what the ISI databases have directly supported and to employ more
sophisticated methods
Physiological roles and regulation of the cryptic prophage-encoded small protein DicB and small RNA DicF in Escherichia coli
No full textBacterial adaptation to stress conditions is vital for their survival in different environments. In the past decade, the important roles that small RNAs (sRNAs) and small proteins perform in regulating the stress response in bacteria has become evident. While the roles of numerous sRNAs and small proteins encoded on the bacterial core genome have been characterized, very few that are phage or prophage-encoded have been studied. This dissertation describes the roles and regulation of the sRNA DicF and small protein DicB encoded on the Qin cryptic prophage of Escherichia coli K12. Bacterial genomes harbor cryptic prophages that have lost genes required for induction, excision from host chromosomes, or production of phage progeny. DicB and DicF are encoded on the dicBF operon, which is located in the immunity region of the cryptic prophage Qin. DicB and DicF have previously been implicated in inhibiting cell division of the host bacterium. In this study, we show that the small protein DicB protects the host cells from phage infection. DicB specifically inhibits infection by λ and other phages that use ManYZ inner membrane proteins to inject their DNA into the host cell. DicB also inhibits the canonical function of ManYZ, which is mannose sugar transport. We demonstrated that the previously known interaction between DicB and MinC, a host protein involved in proper positioning of the Z ring during cell division, is necessary for the DicB-dependent phenotypes involving ManYZ identified in this study. The sRNA DicF is widely conserved is many E. coli strains and inhibits ftsZ mRNA translation in E. coli. In this study, we establish the mechanism of this regulation by characterizing the base pairing interaction between DicF and ftsZ, and delineate the roles of other host factors involved in this regulation. Additionally, we identify new mRNA targets of the sRNA DicF, that are primarily involved in host cell metabolism. The final part of this thesis work describes the genetic mechanisms defining the regulation of the dicBF operon. The dicBF operon is constantly repressed under laboratory conditions by the repressor DicA. In this work, we identified that the protein Rem impairs DicA repression of the dicBF operon and is the antirepressor of DicA. We show that Rem induces expression of dicB and dicF, which leads to the concomitant cell filamentation phenotype as DicB and DicF are cell division inhibitors. In the absence of the dicBF gene products, the promoter of the dicBF operon was found to undergo spontaneous induction in a subset of cells, reminiscent of the λ phage bistable genetic switch. Lastly, urea and high temperature were identified as strong inducers of the dicBF promoter in strains deleted for the dicBF operon. Our results suggest that expression of the dicBF operon is regulated in multiple ways in E. coli K12, indicative of the complex relationship that exists between the host cell and cryptic prophages.Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2023-05-01The student, Preethi Narayani Thattai Ragunathan, accepted the attached license on 2020-12-22 at 15:01.The student, Preethi Narayani Thattai Ragunathan, submitted this Dissertation for approval on 2020-12-22 at 16:38.This Dissertation was approved for publication on 2020-12-23 at 13:10.DSpace SAF Submission Ingestion Package generated from Vireo submission #16136 on 2021-09-16 at 17:01:29Made available in DSpace on 2021-09-17T02:34:10Z (GMT). No. of bitstreams: 2
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Lift date: 2023-09-17T02:34:57Z
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Dr. Glendon Swarthout
No full textHosted by Roger M. Busfield, MSU Assistant Professor of Speech and Theater, Meet the Author is designed to introduce a general audience to a contemporary author and their work through in-depth interviews. This episode features a conversation between Dr. Glendon Swarthout, prolific author and English professor at MSU, and assistant professors Sam S. Baskett and Theodore B. Strandness
Simulation of thermal plant optimization and hydraulic aspects of thermal distribution loops for large campuses
No full textFollowing an introduction, the author describes Texas A&M University and its utilities system. After that, the author presents how to construct simulation models for chilled water and heating hot water distribution systems. The simulation model was used in a $2.3 million Ross Street chilled water pipe replacement project at Texas A&M University. A second project conducted at the University of Texas at San Antonio was used as an example to demonstrate how to identify and design an optimal distribution system by using a simulation model. The author found that the minor losses of these closed loop thermal distribution systems are significantly higher than potable water distribution systems. In the second part of the report, the author presents the latest development of software called the Plant Optimization Program, which can simulate cogeneration plant operation, estimate its operation cost and provide optimized operation suggestions. The author also developed detailed simulation models for a gas turbine and heat recovery steam generator and identified significant potential savings. Finally, the author also used a steam turbine as an example to present a multi-regression method on constructing simulation models by using basic statistics and optimization algorithms. This report presents a survey of the author??s working experience at the Energy Systems Laboratory (ESL) at Texas A&M University during the period of January 2002 through March 2004. The purpose of the above work was to allow the author to become familiar with the practice of engineering. The result is that the author knows how to complete a project from start to finish and understands how both technical and nontechnical aspects of a project need to be considered in order to ensure a quality deliverable and bring a project to successful completion. This report concludes that the objectives of the internship were successfully accomplished and that the requirements for the degree of Degree of Engineering have been satisfied
Intern experience at CH���M Hill, Inc.: an internship report
No full textIncludes author's vita"Submitted to the College of Engineering of Texas A&M University in partial
fulfillment of the requirement for the degree of Doctor of Engineering."Includes bibliographical referencesA review of the author's internship experience with CH���M HILL, Inc.
during the period September 1975 through May 1976 is presented. During this nine month
internship the author worked as an Engineer II in the Industrial Processes discipline of this
large consulting engineering firm... The author's prime responsibility was as one of three
lead design engineers on the design of a large wastewater treatment facility for a pulp mill
in Hoquiam, Washington owned by ITT Rayonier Inc. The work generally consisted of the design
of individual treatment units and associated piping and pumping. The purpose of the project
was to provide wastewater treatment capabilities that would satisfy the effluent limitations
(standards) imposed upon the mill by the State of Washington Department of Ecology and the
U.S. Environmental Protection Agency. The author's assignment also entailed necessary
interaction with the project manager and other CH���M HILL design engineers and support staff
members, the client's representatives, and representatives of two other consulting engineering
firms working on the project. Thus, the internship position at CH���M HILL provided considerable
experience coordinating the author's work with the work of other engineers, guiding the design
and administrative efforts of a support staff, and interacting regularly with the client and
other consulting firms. This broad exposure to a variety of engineering and organizational
problems provided a valuable educational experience
Transition to turbulence in a qblique shock-wave/boundary-layer interaction at M=15
No full textDirect numerical simulations are carried out for different forcing techniques to trigger transition during the interaction between an oblique shock-wave and a laminar boundary-layer at M = 1.5. Three forcing methods are used: a) forcing of oblique unstable modes, whose shape and behaviour are determined by the local linear stability theory, b) broadband free-stream acoustic disturbances, and c) a cold plasma flow control device. While the oblique-mode breakdown is dominant for low-amplitude forcing, long streaky structures drive the transition process in a high-amplitude disturbance environment. LES are also performed on the experimental setup by the Institute of Theoretical and Applied Mechanics (ITAM) from Novosibirsk State University with cold plasma actuation. As well as the disturbance type, the effect of Reynolds number and forcing amplitude will be investigated
Letter from Cedrick M. Shimo to the Office of Redress Administration, June 4, 1991
No full textA letter from Cedrick M. Shimo to the Office of Redress Administration arguing that John Y. Udaka is entitled to a redress payment.These materials are from box 73 and 74 of the Frank Chin Papers. The Frank Chin Papers contain personal and professional correspondence between Frank Chin and Michi Weglyn relating to particular projects on which either author was working as well as files related to the Day of Remembrance Tribute to Michi Weglyn
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