Title Page
Contents
Abstract 11
I. INTRODUCTION 14
II. MATERIALS AND METHODS 23
1. Bioinformatics analysis 23
2. KGN cell culture 23
3. Transfection of KGN cells with a miR-329-3p mimic and inhibitor 24
4. Total RNA extraction and quantitative real-time PCR (qRT-PCR) with KGN cells 24
5. Western blot analysis 26
6. Measurement of E2 levels 26
7. Patients 27
8. Collection of patient's sera, cumulus cells and granulosa cells 28
9. Human serum microRNA extraction and qRT-PCR analysis 29
10. Statistical analysis 30
III. RESULTS 31
1. miR-329-3p expression changes in POR in vitro model 31
2. Bioinformatics analysis to identify miRNA target genes 33
3. miR-329-3p regulates FSH signaling pathway by targeting ADCY9 and PRKACB in KGN cells 37
4. miR-329-3p regulates steroidogenesis in KGN cells 40
5. The clinical characteristics of Control and POR patients 42
6. FSHR and aromatase expression in granulosa cells of control and POR patients 45
7. miR-329-3p expression is elevated in the sera, granulosa cells of POR patients 45
IV. DISCUSSION 49
V. CONCLUSION 56
VI. REFERENCES 59
ABSTRACT IN KOREAN 66
Table 1. KEGG pathway analysis of miR-329-3p associated with ovarian... 35
Table 2. Clinical characteristics of Control and POR patients 44
Table 3. List of articles and proposed functions related to miR-329-3p 53
Figure 1. Controlled ovarian hyperstimulation compared to natural... 15
Figure 2. Start of the study: FSHR dysfunction is the main cause... 17
Figure 3. The basic mechanism of miRNA processing pathway 19
Figure 4. Previous study in our laboratory 22
Figure 5. The expression level of miR-329-3p is up-regulated in... 32
Figure 6. The Bioinformatics analysis of selecting miR-329-3p... 34
Figure 7. KEGG pathway analysis: ovarian steroidogenesis pathway in... 36
Figure 8. ADCY9 and PRKACB were predicted to be target genes of... 39
Figure 9. miR-329-3p regulates aromatase protein level and inhibits... 41
Figure 10. The sampling schedule of IVF patients and Quantification of... 47
Figure 11. The expression level of miR-329-3p is elevated in IVF... 48
Figure 12. Adenylyl cyclase and protein kinase A function in FSH... 51
Figure 13. The predicted molecular mechanism of miR-329-3p on... 57
Figure 14. Conclusion 58