FKBP51影响急性淋巴细胞白血病细胞增殖和化疗敏感性
发布时间：2020-12-26 17:00
简介背景:急性淋巴细胞白血病是青少年中最常见的血液系统恶性肿瘤,成人也可发病。耐药是急性淋巴细胞白血病治疗失败的重要原因,临床上迫切需要开发新的治疗方法或靶向治疗药物。FKBP51是亲免素蛋白家族成员,在淋巴细胞生理性表达。FKBP51参与维持细胞生长、细胞恶变以及化疗耐药。目的:利用Jurkat细胞系,研究FKBP51蛋白对急性淋巴细胞白血病细胞生长和化疗药物敏感性的影响。探讨FKBP51作为急性淋巴细胞白血病发展/演化和对抗癌药物耐药的生物学标志的可能性。方法:用慢病毒载体包装FKBP51,感染Jurkat细胞,建立过表达FKBP51的Jurkat细胞系,用shRNA-FKBP51质粒转染Jurkat细胞,建立敲除FKBP51的Jurkat细胞系。western blot检测FKBP51蛋白表达水平,验证细胞建系成功。经CCK-8分析检测细胞增殖,Muse（?）细胞分析仪检测细胞周期分布,western blot研究丝氨酸/苏氨酸蛋白激酶（AKT）和核因子κB（NF-κB）信号通路。统计学处理:应用SPSS 25.0和Graph Pad Prism 6统计软件进行统计分析,实验数据...
【文章来源】：山东大学山东省 211工程院校 985工程院校 教育部直属院校
【文章页数】：120 页
【学位级别】：博士
【文章目录】：
Abstract
中文摘要
附录
Thesis framework
Chapter One:General Introduction
1.1 Statement
1.2 Hypothesis
1.3 Aim
1.4 Obectives
Chapter Two:Literature Review
2.1 Hematopoiesis
2.2 Cytokines
2.2.1 Functional classification
2.2.2 Role of Endogenous Cytokines in Health and Disease
2.2.3 Application of Therapeutic drugs
2.3 Cancer
2.3.1 History of Cancer
2.3.2 The hallmarks of Cancer
2.4 Leukemia
2.4.1 Acute lymphoblastic leukemia
2.4.1.1 Epidemiology
2.4.1.2 Risk Factors
2.4.1.3 Pathogenesis
2.4.1.4 Classification of Acute lymphoblastic leukemia
2.5 Philadelphia chromosome （Ph）
2.5.1 Leukemogenesis-How?
2.5.2 Breakpoints of translocation t （9;22） BCR-ABL translocation
2.6 FK506 binding protein 51
2.6.1 Functions of the FKBP51
2.6.2 Clinical Significance
2.6.2.1 Expression of FK506 binding protein51 in Cancer
2.7 A serine /threonine- protein kinase
2.7.1 Pathophysiology of AKT
2.7.2 AKT and FKBP51
2.8. Nuclear factor kappa- light-chain- enhancer of activated B cell
2.8.1 NF-κB signaling pathways
2.8.2 NF-κB and FKBP51
2.9 Chemotherapy
2.9.1 Treatment of Chemotherapy is divers from other theray
2.9.2 Cell cycle phases
2.10 Dexamethasone
2.11 Lipopolysaccharide
2.12 Tumor necrosis factor-alpha Tumor necrosis factor-alpha
2.12.1 Role of TNF-α
2.13 Jurkat Cell line
2.14 Lentiviiral Vectors
2.14.1 Factors Influencing the Infectivity of Lentiviral Vectors
Chapter Three: Materials and methods
3.1 Materials
3.1.1 Cell Cultures and treatments
3.2 Common related materials
3.3 Cell Cultures
3.3.1 Principle
3.3.2 Method
3.3.3 Multiplicity of Infection （MOI）
3.4 RNA Isolation and Real-Time Quantitative RT-PCR
3.4.1 Principle
3.4.2 Method
3.5 Cell proliferation assay
3.5.1 Principle
3.5.2 Method
3.6 Cell Cycle assay
3.6.1 Principle
3.6.2 Method
3.7 Western blot assay
3.7.1 Principle
3.7.2 Method
3.8 FKBP51-shRNA （Knockdown）
3.8.1 Principle
3.8.2 Method
3.9 FKBP51 Regulated AKT Pathway
3.9.1 Principle
3.9.2 Method
3.10 NF-κB Signaling pathway
3.10.1 Principle
3.10.2 Method
3.11 Chemosensitivity Assay
3.11.1 Principle
3.11.2 Method
3.12 Statistical analysis
Chapter Four- Results
4.1 The Efficiency of Infection
4.2 Overexpression of FKBP51 in ALL Cell lines
4.2.1 Western blotting
4.2.2 Cell proliferation/viability assay
4.3 The shRNA- FKBP51 （Knockdown）
4.4 The role of FKBP51 in AKT Signaling Pathway
4.5 Western bloting analysis of FKBP51 overexpression and NF-κB kinaseactivity
4.6 Western bloting analysis of shRNA-FKBP51 and NF-κB kinase activity
4.7 CCK-8 assay, negative control and transfected Jurkat cell lines
Chapter Five: Discussion
Chapter Six: Conclusion and future perspective
6.1 Conclusion
6.2 Future perspective
Chapter Seven:References
Thesis Part Two Effect of Helicobacter pylori Infection on Hematological Parameters in KostiSudan
附录
Chapter One: Intoduction
1.1 Statement
1.2 Hypothesis
1.3 Obiective
1.4 Rationale
Chapter Two:Literature review
2.1 Helicobacter pylori infection
2.1.1 Prevalence of Helicobacter pylori infection
2.1.2. Causes of H. pylori
2.1.3 Pathophysiology
2.2. H. pylori and Iron Deficiency Anemia
2.3 Lactoferrin
2.4 H. pylori and Megaloblastic Anemia
12 deficiency"> 2.4.1 Causes of vitamin B12 deficiency
2.5 Pernicious anemia
2.5.1 Schilling test
2.6 Gastrointestinal Tract Endoscopy
2.7 H.pylori and CagA
2.8 H.pylori and signaling pathways
2.9 Tumor suppressor p53
2.10 Wnt signaling pathways
2.11 H. pylori and obesity
2.12 Diagnostic Methods of H. Pylori
Chapter Three: Materials and Methods
3.1 Study population and design
3.2 Inclusion and Exclusion criteria
3.3 Sampling procedure
3.4 Estimation of hematological parameters
3.4.1 Principle
3.4.2 Method
3.5 Reticulocytes （Retic） Count
3.5.1 Principle
3.5.2 Method
3.5.3. Interpretation of the results
3.5.3.1 Normal account
3.5.3.2 Decreased reticulocyte count
3.5.3.4 Increased reticulocyte count
3.6 H. pylori screening
3.7 Statistical analysis
3.8. Ethical statement
Chapter Four: Results
4.1 Comparison of Demographic and Hematological Parameters between Patientsand Control Groups
4.2 Evaluations of Hematological Parameters between Patients, Control groupsand their correlation with H.pylori
4.3 Platelet count between control and patients'groups
4.4 Morphological examination of reticulocyte count
4.5 Morphological blood picture
Chapter Five:Discssion
Chapter Six:Conclusion and future perspective
6.1 Conclusion
6.2 future perspective
Chapter Seven: References
Chapter Eight: Appendices
Appendix 1-Information and Consent Form
Appendix 2- Questionnaire
Appendix 3- The formula to calculate the Reticulocytes count
Appendix 4- Red blood cell indices, the formula for calculation
Appendix 5- Buffers and solutions
Appendix 6- Equipment
Appendix 7- Software
Appendix 8- cell cycle Phases
Dedication
Acknowledgements
List of Publications
Published work
附件
【参考文献】：
期刊论文
[1]Aberrant control of NF-κB in cancer permits transcriptional and phenotypic plasticity, to curtail dependence on host tissue: molecular mode[J]. Spiros A.Vlahopoulos. Cancer Biology & Medicine. 2017(03)
[2]Therapeutic upper gastrointestinal tract endoscopy in Paediatric Gastroenterology[J]. Imdadur Rahman,Praful Patel,Philip Boger,Shahnawaz Rasheed,Mike Thomson,Nadeem Ahmad Afzal. World Journal of Gastrointestinal Endoscopy. 2015(03)
[3]Beyond the stomach: An updated view of Helicobacter pylori pathogenesis, diagnosis, and treatment[J]. Traci L Testerman,James Morris. World Journal of Gastroenterology. 2014(36)
[4]Extraintestinal manifestations of Helicobacter pylori:A concise review[J]. Frank Wong,Erin Rayner-Hartley,Michael F Byrne. World Journal of Gastroenterology. 2014(34)
[5]Classification of anemia for gastroenterologists[J]. Jose Antonio Moreno Chulilla,Maria Soledad Romero Colás,Martín Gutiérrez Martín. World Journal of Gastroenterology. 2009(37)
[6]Helicobacter pylori and other Helicobacter species DNA in human bile samples from patients with various hepato-biliary diseases[J]. Santosh K Tiwari,Aleem A Khan,Mohd Ibrahim,Mohd Aejaz Habeeb,C M Habibullah. World Journal of Gastroenterology. 2006(14)
[7]Wnt signaling in disease and in development[J]. Roel NUSSE. Cell Research. 2005(01)
[8]OVEREXPRESSION OF Akt-1 GENE IN HUMAN HEPATOCELLULAR CARCINOMA[J]. 刘连新,刘芝华,姜洪池,綦书抑,张伟辉,朱安龙,王秀琴,吴旻. Chinese Journal of Cancer Research. 2002(03)
本文编号：2940106
【文章来源】：山东大学山东省 211工程院校 985工程院校 教育部直属院校
【文章页数】：120 页
【学位级别】：博士
【文章目录】：
Abstract
中文摘要
附录
Thesis framework
Chapter One:General Introduction
1.1 Statement
1.2 Hypothesis
1.3 Aim
1.4 Obectives
Chapter Two:Literature Review
2.1 Hematopoiesis
2.2 Cytokines
2.2.1 Functional classification
2.2.2 Role of Endogenous Cytokines in Health and Disease
2.2.3 Application of Therapeutic drugs
2.3 Cancer
2.3.1 History of Cancer
2.3.2 The hallmarks of Cancer
2.4 Leukemia
2.4.1 Acute lymphoblastic leukemia
2.4.1.1 Epidemiology
2.4.1.2 Risk Factors
2.4.1.3 Pathogenesis
2.4.1.4 Classification of Acute lymphoblastic leukemia
2.5 Philadelphia chromosome （Ph）
2.5.1 Leukemogenesis-How?
2.5.2 Breakpoints of translocation t （9;22） BCR-ABL translocation
2.6 FK506 binding protein 51
2.6.1 Functions of the FKBP51
2.6.2 Clinical Significance
2.6.2.1 Expression of FK506 binding protein51 in Cancer
2.7 A serine /threonine- protein kinase
2.7.1 Pathophysiology of AKT
2.7.2 AKT and FKBP51
2.8. Nuclear factor kappa- light-chain- enhancer of activated B cell
2.8.1 NF-κB signaling pathways
2.8.2 NF-κB and FKBP51
2.9 Chemotherapy
2.9.1 Treatment of Chemotherapy is divers from other theray
2.9.2 Cell cycle phases
2.10 Dexamethasone
2.11 Lipopolysaccharide
2.12 Tumor necrosis factor-alpha Tumor necrosis factor-alpha
2.12.1 Role of TNF-α
2.13 Jurkat Cell line
2.14 Lentiviiral Vectors
2.14.1 Factors Influencing the Infectivity of Lentiviral Vectors
Chapter Three: Materials and methods
3.1 Materials
3.1.1 Cell Cultures and treatments
3.2 Common related materials
3.3 Cell Cultures
3.3.1 Principle
3.3.2 Method
3.3.3 Multiplicity of Infection （MOI）
3.4 RNA Isolation and Real-Time Quantitative RT-PCR
3.4.1 Principle
3.4.2 Method
3.5 Cell proliferation assay
3.5.1 Principle
3.5.2 Method
3.6 Cell Cycle assay
3.6.1 Principle
3.6.2 Method
3.7 Western blot assay
3.7.1 Principle
3.7.2 Method
3.8 FKBP51-shRNA （Knockdown）
3.8.1 Principle
3.8.2 Method
3.9 FKBP51 Regulated AKT Pathway
3.9.1 Principle
3.9.2 Method
3.10 NF-κB Signaling pathway
3.10.1 Principle
3.10.2 Method
3.11 Chemosensitivity Assay
3.11.1 Principle
3.11.2 Method
3.12 Statistical analysis
Chapter Four- Results
4.1 The Efficiency of Infection
4.2 Overexpression of FKBP51 in ALL Cell lines
4.2.1 Western blotting
4.2.2 Cell proliferation/viability assay
4.3 The shRNA- FKBP51 （Knockdown）
4.4 The role of FKBP51 in AKT Signaling Pathway
4.5 Western bloting analysis of FKBP51 overexpression and NF-κB kinaseactivity
4.6 Western bloting analysis of shRNA-FKBP51 and NF-κB kinase activity
4.7 CCK-8 assay, negative control and transfected Jurkat cell lines
Chapter Five: Discussion
Chapter Six: Conclusion and future perspective
6.1 Conclusion
6.2 Future perspective
Chapter Seven:References
Thesis Part Two Effect of Helicobacter pylori Infection on Hematological Parameters in KostiSudan
附录
Chapter One: Intoduction
1.1 Statement
1.2 Hypothesis
1.3 Obiective
1.4 Rationale
Chapter Two:Literature review
2.1 Helicobacter pylori infection
2.1.1 Prevalence of Helicobacter pylori infection
2.1.2. Causes of H. pylori
2.1.3 Pathophysiology
2.2. H. pylori and Iron Deficiency Anemia
2.3 Lactoferrin
2.4 H. pylori and Megaloblastic Anemia
12 deficiency"> 2.4.1 Causes of vitamin B12 deficiency
2.5 Pernicious anemia
2.5.1 Schilling test
2.6 Gastrointestinal Tract Endoscopy
2.7 H.pylori and CagA
2.8 H.pylori and signaling pathways
2.9 Tumor suppressor p53
2.10 Wnt signaling pathways
2.11 H. pylori and obesity
2.12 Diagnostic Methods of H. Pylori
Chapter Three: Materials and Methods
3.1 Study population and design
3.2 Inclusion and Exclusion criteria
3.3 Sampling procedure
3.4 Estimation of hematological parameters
3.4.1 Principle
3.4.2 Method
3.5 Reticulocytes （Retic） Count
3.5.1 Principle
3.5.2 Method
3.5.3. Interpretation of the results
3.5.3.1 Normal account
3.5.3.2 Decreased reticulocyte count
3.5.3.4 Increased reticulocyte count
3.6 H. pylori screening
3.7 Statistical analysis
3.8. Ethical statement
Chapter Four: Results
4.1 Comparison of Demographic and Hematological Parameters between Patientsand Control Groups
4.2 Evaluations of Hematological Parameters between Patients, Control groupsand their correlation with H.pylori
4.3 Platelet count between control and patients'groups
4.4 Morphological examination of reticulocyte count
4.5 Morphological blood picture
Chapter Five:Discssion
Chapter Six:Conclusion and future perspective
6.1 Conclusion
6.2 future perspective
Chapter Seven: References
Chapter Eight: Appendices
Appendix 1-Information and Consent Form
Appendix 2- Questionnaire
Appendix 3- The formula to calculate the Reticulocytes count
Appendix 4- Red blood cell indices, the formula for calculation
Appendix 5- Buffers and solutions
Appendix 6- Equipment
Appendix 7- Software
Appendix 8- cell cycle Phases
Dedication
Acknowledgements
List of Publications
Published work
附件
【参考文献】：
期刊论文
[1]Aberrant control of NF-κB in cancer permits transcriptional and phenotypic plasticity, to curtail dependence on host tissue: molecular mode[J]. Spiros A.Vlahopoulos. Cancer Biology & Medicine. 2017(03)
[2]Therapeutic upper gastrointestinal tract endoscopy in Paediatric Gastroenterology[J]. Imdadur Rahman,Praful Patel,Philip Boger,Shahnawaz Rasheed,Mike Thomson,Nadeem Ahmad Afzal. World Journal of Gastrointestinal Endoscopy. 2015(03)
[3]Beyond the stomach: An updated view of Helicobacter pylori pathogenesis, diagnosis, and treatment[J]. Traci L Testerman,James Morris. World Journal of Gastroenterology. 2014(36)
[4]Extraintestinal manifestations of Helicobacter pylori:A concise review[J]. Frank Wong,Erin Rayner-Hartley,Michael F Byrne. World Journal of Gastroenterology. 2014(34)
[5]Classification of anemia for gastroenterologists[J]. Jose Antonio Moreno Chulilla,Maria Soledad Romero Colás,Martín Gutiérrez Martín. World Journal of Gastroenterology. 2009(37)
[6]Helicobacter pylori and other Helicobacter species DNA in human bile samples from patients with various hepato-biliary diseases[J]. Santosh K Tiwari,Aleem A Khan,Mohd Ibrahim,Mohd Aejaz Habeeb,C M Habibullah. World Journal of Gastroenterology. 2006(14)
[7]Wnt signaling in disease and in development[J]. Roel NUSSE. Cell Research. 2005(01)
[8]OVEREXPRESSION OF Akt-1 GENE IN HUMAN HEPATOCELLULAR CARCINOMA[J]. 刘连新,刘芝华,姜洪池,綦书抑,张伟辉,朱安龙,王秀琴,吴旻. Chinese Journal of Cancer Research. 2002(03)
本文编号：2940106
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