Title Page
Contents
Ⅰ. Abstract 9
Ⅱ. Introduction 11
Ⅲ. Materials and Methods 15
Reagents 15
Cell culture 15
MKP-3 knockout (KO) clone establishment 15
Western blotting 16
RNA sequencing 16
CCK-8 assay 16
Mouse Apoptosis Signaling Pathway Array 17
Intracellular reactive oxygen species (ROS) 17
RNA isolation, reverse transcription, and quantitative reverse-transcription PCR 18
Senescence-associated b-galactosidase (SA-β-Gal) staining 18
Mouse colon cancer cells and macrophage co-culture 19
Mouse cytokine array panel A 19
Mouse VEGFA ELISA assay 19
General PGE2 / Prostaglandin E2 ELISA assay 20
Mouse high mobility group protein B1 (HMGB1) ELISA assay 20
Mouse LDL R immunoassay 20
Statistics 21
Ⅳ. Results 24
Loss of MKP-3 affects cancer cells by altering the MAPK and STAT pathways 24
Gene expression in MKP-3 KO cells 26
The loss of MKP-3 influences the resistance of cancer cells to chemotherapy and apoptotic pathway 29
MKP-3 counteracts oxidative stress by regulating ROS levels 32
Loss of MKP-3 accelerates cellular senescence 36
Loss of MKP-3 induces M2 polarization 40
Cytokine profiling in MKP-3 KO 42
Loss of MKP-3 does not affect glucose metabolism in cancer cells 45
MKP-3 does not regulate EMT at the basal level 47
Ⅴ. Discussion 49
Reference 52
국문 초록 61
Table 1. List of primary antibodies 22
Table 2. List of primer sequences for real-time quantitative PCR 23
Figure 1. Loss of MKP-3 affects cancer cells by altering the MAPK and STAT signaling pathways 25
Figure 2. The expression of well-known cancer-related genes did not differ between WT and MKP-3 KO cells 28
Figure 3. MKP-3 KO cells show reduced sensitivity to doxorubicin (DOXO) 31
Figure 4. MKP-3 regulates NOX4 and NQO1 to inhibit the ROS-generating entry process 35
Figure 5. Loss of MKP-3 accelerates aging by overactivation of the ERK Pathway 39
Figure 6. MKP-3 recruits macrophages, creating a more favorable environment for cancer cells 41
Figure 7. The effects of MKP-3 on cytokine levels require further study 44
Figure 8. MKP-3 does not affect glucose metabolism in cancer cells 46
Figure 9. MKP-3 does not affect EMT at the basal levels 48