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Role of BTG2/TIS21/PC3 in differentiation of myeloid leukemia cells and its regulation under stress conditions
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 임인경 | - |
| dc.contributor.author | MUHAMMAD IMRAN | - |
| dc.date.accessioned | 2019-10-21T07:23:43Z | - |
| dc.date.available | 2019-10-21T07:23:43Z | - |
| dc.date.issued | 2014-02 | - |
| dc.identifier.other | 16480 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/18508 | - |
| dc.description | 학위논문(박사)--아주대학교 일반대학원 :의생명학과,2014. 2 | - |
| dc.description.tableofcontents | ABSTRACT ························································································ i TABLE OF CONTENTS ········································································ iv LIST OF FIGURES ············································································ viii Part 1 I. INTRODUCTION ··············································································· 1 II. MATERIALS AND METHODS ····························································· 7 1. Cell culture ······················································································· 7 2. Adenoviral transduction of HL-60 cells with TIS21 gene ································· 7 3. Differentiation analyses ········································································ 7 4. RNA extraction, semiquantitative RT-PCR and real time PCR ··························· 8 5. RNA Interference ·············································································· 10 6. Immunoblot and immunoprecipitation analyses ············································ 10 7. Statistical Analyses ············································································ 10 III. RESULTS ····················································································· 11 A. ATRA-induces granulocytic differentiation of HL-60 cells ······························ 11 B. ATRA upregulates Btg2 expression in HL-60 cells ······································· 14 C. TIS21 enhances ATRA-induced differentiation of HL-60 cells ························· 16 D. TIS21 enhances ATRA-induced differentiation via down-regulation of c-Myc in HL-60 cell ·························································································· 19 E. TIS21 decreases stability of c-Myc protein in response to ATRA treatment ··········· 25 F. Transduction of shTIS21 abrogates TIS21 effect on c-Myc expression and HL-60 cells differentiation ··············································································· 27 v G. ATRA plus TIS21 increased Erk1/2 activity, but inhibited Akt with subsequent GSK-3β activation ················································································ 29 H. TIS21 enhanced downregulation of c-Myc by activating GSK-3β ······················ 36 I. TIS21 enhanced ATRA-mediated c-Myc degradation in the proteosome ··············· 39 IV. DISCUSSION················································································· 42 V. CONCLUSION ················································································ 45 Part 2 I. INTRODUCTION ·············································································· 46 II. MATERIALS AND METHODS ···························································· 50 1. Cell culture ······················································································ 50 2. RNA extraction and semi quantitative RT-PCR ············································ 50 3. Immunoblot analysis ·········································································· 52 4. Cells fractionation ·············································································· 52 5. Measurement of intracellular ROS level and cell cycle analysis ························· 52 6. Chromatin immunoprecipitation (ChIP) assay ············································· 53 7. Cells Proliferation Assay ······································································ 53 8. Transfection of PKC- ········································································ 8. Statistical analysis ············································································· 54 III. RESULTS ····················································································· 55 A. Btg2 is upregulated under serum deprivation ·············································· 55 B. Serum deprivation-induced reactive oxygen species generation upregulates Btg2 expression ··················································································· 60 C. Serum deprivation induces NF-B activation ·············································· 63 D. NF-B regulates Btg2 expression under serum deprivation ······························ 66 vi E. SP1 and DNA damage signals do not regulate Btg2 expression under serum deprivation ························································································· 69 F. Exogenous H2O2 regulates Btg2 expression via NF-B activation in DLD-1 cells ···· 71 G. Doxorubicin induces Btg2 expression via ROS-NF-B pathway ······················· 73 H. ROS regulate NF-B activation along with Btg2 expression via PKC activity ······· 76 I. PKC- regulates NF-B activity and Btg2 expression in ROS dependent manner ···· 81 J. ROS regulate NFB-Btg2 under serum deprivation via MAPK pathway activation ·· 83 K. Btg2 reduces cells proliferation expression under serum deprivation, H2O2 and Doxo treatment ···················································································· 86 IV. DISCUSSION················································································· 89 V. CONCLUSION ················································································ 93 Part 3 I. INTRODUCTION ·············································································· 94 II. MATERIALS AND METHODS ···························································· 97 1. Cell culture ······················································································ 97 2. Differentiation analyses ······································································· 97 3. RNA extraction, semiquantitative RT-PCR and real time PCR ·························· 97 4. Immunoblot and immunoprecipitation analyses ············································ 98 5. Cells fractionation ············································································· 98 6. Measurement of intracellular ROS level ···················································· 99 7. Statistical Analyses ············································································ 99 III. RESULTS ··················································································· 100 A. ATRA induces macrophage differentiation under reduced serum concentration ··· 100 B. ATRA enhances NF-B activation in myeloid leukemia cells ························ 104 vii C. Inhibition of NF-B abrogates ATRA induced macrophage differentiation of HL-60 cells······················································································· 107 D. Activation of NF-B induces macrophage differentiation in response to ATRA Treatment ························································································ 109 E. ATRA enhances NF-B activation and induces macrophage differentiation via MAPK ···························································································· 112 F. ATRA induced C/EBPα expression via MAPK is lower in cells under serum deprivation ······················································································· 115 IV. DISCUSSION··············································································· 118 V. CONCLUSION ·············································································· 121 VI. REFERENCES ············································································· 122 국문 요약 ························································································ 148 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | Role of BTG2/TIS21/PC3 in differentiation of myeloid leukemia cells and its regulation under stress conditions | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.department | 일반대학원 의생명과학과 | - |
| dc.date.awarded | 2014. 2 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 609677 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000016480 | - |
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- Graduate School of Ajou University > Department of Biomedical Sciences > 3. Theses(Master)
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