The regulatory role of mixed lineage kinase 4 beta in MAPK signaling and ovarian cancer cell invasion

The regulatory role of mixed lineage kinase 4 beta in MAPK signaling and ovarian cancer cell invasion
Table of Contents
Abstract
Acknowledgements
Table of Contents
List of Figures
List of Abbreviations
1 Introduction
1.1 The Mitogen-activated protein kinase signaling cascade
1.2 Characteristics and functions of MAPK proteins
1.2.1 The ERK1/2 pathway
1.2.2 The JNK pathway
1.2.3 The p38 pathway
1.2.4 The ERK5 pathway
1.3 The matrix metalloproteinases
1.4 The MAP2Ks
1.5 The MAP3Ks
1.5.1 The MEKK group
1.5.2 The Raf MAP3Ks
1.5.3 The TAK1 MAP3K group
1.5.4 The TAO/Tpl2 and Mos MAP3K groups
1.5.5 The MLK family of MAP3Ks
1.5.5.1 The DLK subgroup
1.5.5.2 The ZAK subgroup
1.6 The MLK subfamily
1.6.1 MLK1 and MLK2
1.6.2 MLK3 activation
1.7 MLK3 signaling
1.7.1 MLK3 signaling in cancer
1.8 MLK4: characteristics and function
1.9 Significance
2 Materials and Methods
2.1 Cell culture
2.2 Expression vectors
2.3 Plasmids and siRNA transfections
2.4 Immunoblotting
2.5 Preparation of whole cell extracts and treatments
2.6 Immunoprecipitation
2.7 MLK4β kinase assay
2.8 Cell proliferation assay
2.9 Luciferase assay
2.10 Invasion assay
2.11 Gelatin zymography
3 Results
3.1 The role of MLK4β in p38 signaling
3.1.1 The effect of ectopic expression of MLK4β on p38 activation
3.1.2 The effect of endogenous MLK4 on the activation of p38.
3.2 The effect of MLK4 on MEK3/MEK6 activation
3.3 The role of MLK4β in NF-κB signaling
3.4 Comparison of the effects of MLK4β and MLK3 on p38 activation
3.5 The effects of MLK3 and MLK4 on ERK and JNK activation
3.6 MLK4β is not an upstream activator of MEK4
3.7 MLK4β kinase activity
3.8 The effect of MLK4β on MLK3 activation
3.9 The correlation between MLK4β expression and active MLK3 in different cell lines
3.10 MLK4β associates with MLK3
3.11 The effect of MLK4 on cell proliferation
3.12 MLK3 is required for cell invasion in ovarian cancer cells
3.13 MLK4β inhibits SKOV3 cell invasion
3.14 MLK3 regulates MMP-2 and MMP-9 enzyme activity
3.15 MLK4β reduces MMP-9 activity in SKOV3 cells
4 Discussion
References
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