Endometrial cancer is one of the most significant gynaecological malignancies that affect women from New Zealand and the rest of the world. One of the critical stages in the development of a tumour is the onset of hypoxia. The malignancy responds by having raised levels of Hypoxia Inducible Factor (HIF) that in turn induces increased production of Vascular Endothelial Growth Factor (VEGF). VEGF is a potent angiogenic factor that will mediate vascular supply of nutrients and oxygen to the developing tumour. The aim of this study was to investigate whether two compounds found in extracts of plant materials, Resveratrol (Resveratrol) and Epigallocatechin gallate (EGCG), altered the levels of VEGF in the supernatant of cultured endometrial cancer cells. Resveratrol is a phytoalexin that is found in many foods, such as grapes, nuts and berries, as well as in high concentrations in some red wines. 100 µM of resveratrol was added to cell cultures for 24 hours. VEGF levels in the supernatant were then analysed using ELISA. Resveratrol was found to have significant inhibitory effects in both primary endometrial cancer cell cultures and immortalised endometrial cancer cell cultures. Resveratrol was also shown to reverse the increase in VEGF caused by the hypoxia mimic cobalt chloride (CoCl₂). Epigallocatechin gallate (EGCG) is an antioxidant catechin extracted from green tea. The effect of EGCG was analysed using the same method as for resveratrol. 100 µM of EGCG was also shown to have a significant inhibitory effect on the level of VEGF in the supernatant of cultured endometrial cancer cells, as well as reducing the effect of CoCl₂. These results suggest that selected food compounds, resveratrol and EGCG, can reduce VEGF levels by inhibiting HIF. Further investigation This may have anti-tumour effects in women with endometrial cancer.

Abstract Aim: It has been suggested that chronic hyperinsulinemia from insulin resistance is involved in the etiology of endometrial cancer (EC). We performed a systematic review and meta-analysis to assess whether insulin resistance is associated with the risk of EC. Methods: We searched PubMed-Medline, Embase, Scopus, and Web of Science for articles published from database inception through 30th September 2014. We included all observational studies evaluating components defining insulin resistance in women with and without EC. Quality of the included studies was assessed by NewcastleeOttawa scale. Randomeffects models and inverse variance method were used to meta-analyze the association between insulin resistance components and EC. Results: Twenty-five studies satisfied our inclusion criteria. Fasting insulin levels (13 studies, n Z 4088) were higher in women with EC (mean difference [MD] 33.94 pmol/L, 95% confi- dence interval [CI] 15.04e52.85, p Z 0.0004). No differences were seen in postmenopausal versus pre- and postmenopausal subgroup analysis. Similarly, non-fasting/fasting C-peptide levels (five studies, n Z 1938) were also higher in women with EC (MD 0.14 nmol/L, 95% CI 0.08e0.21, p < 0.00001). Homeostatic model assessment - insulin resistance (HOMA-IR) values (six studies, n Z 1859) in EC patients were significantly higher than in women without EC (MD 1.13, 95% CI 0.20e2.06, p Z 0.02). There was moderate-to-high heterogeneity among the included studies. Conclusion: Currently available epidemiologic evidence is suggestive of significantly higher risk of EC in women with high fasting insulin, non-fasting/fasting C-peptide and HOMAIR values.

MSc (Med), Biostatistics and Epidemiology, Faculty of Health Sciences, University of the Witwatersrand, 2009 / Introduction: Endometrial cancer is a gynaecological cancer that mostly affects women in their sixth and seventh decades of life. It is the fourth most common malignancy in women and ranks eighth among all causes of female cancer in terms of age-adjusted mortality. In developed and numerous developing countries endometrial cancer, as well as other types of cancer in women, is an ever-increasing threat that may be explained, among other reasons, by increased life expectancy and a reduction in fertility or birth rates. Conversely, in South Africa and most other African countries, the previous reasons do not exist, because there is a decline in life expectancy due to increased HIV, low income, and a high fertility rate. International epidemiological studies have established significant relationships between endometrial cancer and risk factors such as the woman’s age, race, early menarche and late menopause, parity, a history of breast or ovarian cancer, the use of endogenous estrogens, concomitant diabetes, family history of breast and ovarian cancer, estrogen therapy, obesity, and the use of tamoxifen. The aim of the study was to identify risk factors associated with endometrial cancer among black South African women. Method: The present case control study comprised black South African women diagnosed with a cancer in Johannesburg, between 1995 and 2005. The study included 592 women aged 27 to 90 years who were admitted to three main public hospitals in the city of Johannesburg with histologically confirmed cancers. 148 cases with endometrial cancer and 444 women with other forms of cancer were analysed. Only newly occurring cases (incident) were included. Women in the control group consisted of those with V cancers not associated with reproductive or hormonal factors, i.e. not cancers of the breast or the female reproductive system. Data handling, cleaning and analysis were done using Stata 9 (STATA). Results: Univariate analysis showed that the risk for endometrial cancer was significantly (P<0.05) affected by: miscarriages, the place of former residence, place of current residence, the use of snuff, wine consumption, age of the youngest child, diabetes, age of menarche, age of menopause, and menstrual status. Smoking was found to be a protective factor for endometrial cancer compared to other cancers. After multivariate adjustment, endometrial cancer risk was significantly (P<0, 05) associated with miscarriages, age at menarche, and earlier completion of childbearing. Smoking remained a protective factor against endometrial cancer. Conclusion: The current study reports similar results to those observed in other international investigations. The risk of endometrial cancer was higher among women who were older, women who experienced miscarriages, and those who fell pregnant early in their reproductive lives. Smoking was a protective factor against endometrial cancer compared to other cancers. However, comparing the cases of endometrial cancer with smoking-associated cancer controls (i.e. lung cancer, oesophageal cancer, and mouth cancer) might have distorted the results. A more appropriate control group for confirming the relationship between smoking and endometrial cancer would be subjects with no cancer. Also, it will be important to evaluate the risk factors for cancer among the other race groups in South Africa.

Ip Toi Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 81-105). / Abstracts in English and Chinese. / CONTENTS --- p.i-iii / ACKNOWLEDGEMENT --- p.iv / ABSTRACT --- p.v-vi / Chapter Chapter One --- INTRODUCTION --- p.1-2 / Chapter Chapter Two --- LITERATURE REVIEW --- p.3-37 / Chapter 2.1 --- Epidemiology and Etiology of Cervical and Endometrial Cancers --- p.3-4 / Chapter 2.1.1 --- Epidemiology and Etiology of Cervical cancer --- p.4 / Chapter 2.1.1.1 --- Incidence and Mortality --- p.4-6 / Chapter 2.1.1.2 --- Etiology --- p.6-8 / Chapter 2.1.2 --- Epidemiology and Etiology of Endometrial Cancer --- p.9 / Chapter 2.1.2.1 --- Incidence and Mortality --- p.9-11 / Chapter 2.1.2.2 --- Rick Factors --- p.11-14 / Chapter 2.2 --- Pathology of Cervical and Endometrial Cancers --- p.14 / Chapter 2.2.1 --- Pathology of Cervical Cancer --- p.14-15 / Chapter 2.2.1.1 --- Macroscopic Appearance --- p.15 / Chapter 2.2.1.2 --- Histology --- p.15-18 / Chapter 2.2.2 --- Staging of Cervical Cancer --- p.19-21 / Chapter 2.2.3 --- Pathology of Endometrial Cancer --- p.21 / Chapter 2.2.3.1 --- Macroscopic Appearance --- p.22 / Chapter 2.2.3.2 --- Histology --- p.22-24 / Chapter 2.2.4 --- Staging of Endometrial Cancer --- p.24-25 / Chapter 2.2 --- Introduction to Microsatellite Instability (MI) --- p.25 / Chapter 2.3.1 --- DNA structure --- p.25-27 / Chapter 2.3.2 --- Microsatellite --- p.27-28 / Chapter 2.3.3 --- Mismatch Repair (MMR) --- p.28-29 / Chapter 2.3.4 --- Microsatellite Instability (MI) --- p.30-33 / Chapter 2.3.5 --- Microsatellite Instability in various cancers --- p.33-37 / Chapter Chapter Three --- MATERIALS AND METHODS --- p.38-50 / Chapter 3.1 --- Materials --- p.38 / Chapter 3.1.1 --- Patients and Specimens --- p.38-39 / Chapter 3.1.2 --- Chemicals and Reagents --- p.39 / Chapter 3.1.2.1 --- Chemicals --- p.39-40 / Chapter 3.1.2.2 --- Solution --- p.40-41 / Chapter 3.1.2.3 --- Microsatellite Markers --- p.42 / Chapter 3.1.3 --- Major Equipment --- p.43 / Chapter 3.2 --- Methodology --- p.43 / Chapter 3.2.1 --- DNA Extraction --- p.43-45 / Chapter 3.2.2 --- DNA Amplification --- p.45 / Chapter 3.2.2.1 --- End-labeling of Primer --- p.45 / Chapter 3.2.2.2 --- Polymerase Chain Reaction (PCR) --- p.46 / Chapter 3.2.3 --- Electrophoresis of PCR Products and Autoradiography --- p.46-49 / Chapter 3.2.4 --- Determination Of Microsatellite Instability (MI) --- p.49 / Chapter 3.3 --- Statistical Analyses --- p.50 / Chapter Chapter Four --- Result --- p.51-66 / Chapter 4.1 --- Microsatellite Instability in Cervical Cancer --- p.51 / Chapter 4.1.1 --- Prevalence of MI in Cervical Cancer --- p.51 -54 / Chapter 4.1.2 --- MI and Age in Cervical Cancer --- p.55 / Chapter 4.1.3 --- MI and Histological Type in Cervical Cancer --- p.55-56 / Chapter 4.1.4 --- MI and Histologic Grades in Cervical Cancer --- p.56-57 / Chapter 4.1.5 --- MI and Clinical stage in Cervical Cancer --- p.57-58 / Chapter 4.1.6 --- MI and Clinical Status in Cervical Cancer --- p.58-59 / Chapter 4.2 --- Microsatellite Instability in Endometrial Cancer --- p.59 / Chapter 4.2.1 --- Prevalence of MI in Endometrial Cancer --- p.59-62 / Chapter 4.2.2 --- MI and Age Groups in Endometrial Cancer --- p.63 / Chapter 4.2.3 --- MI and Histological Type in Endometrial Cancer --- p.63-64 / Chapter 4.2.4 --- MI and Histologic Grades in Endometrial Cancer --- p.64-65 / Chapter 4.2.5 --- MI and Clinical stage of Endometrial Cancer --- p.65 / Chapter 4.2.6 --- MI and Clinical Status in Endometrial Cancer --- p.66 / Chapter Chapter Five --- Discussion --- p.67-77 / Chapter 5.1 --- MI detection --- p.67-71 / Chapter 5.2 --- MI of Cervical Cancer --- p.71 -74 / Chapter 5.3 --- MI of Endometrial Cancer --- p.74-77 / Chapter Chapter Six --- Conclusions --- p.78-80 / Reference --- p.81-112 / Appendix --- p.113-114

Cheung Ka Wai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 83-93). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Publications --- p.ii / Awards --- p.iii / List of abbreviations --- p.iv / List of figures --- p.vi / List of tables --- p.vii / Abstract in English --- p.viii / Abstract in Chinese --- p.ix / Table of Contents / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- literature Review / Chapter 2.1 --- Anatomy and Physiology of Endometrium --- p.2 / Chapter 2.2 --- Endometrial cancer --- p.5 / Chapter 2.2.1 --- Epidemiology --- p.6 / Chapter 2.2.2 --- Etiologies and Risk Factors --- p.7 / Chapter 2.3 --- Pathology --- p.11 / Chapter 2.3.1 --- Grading of endometrial cancer --- p.14 / Chapter 2.3.2 --- Staging of endometrial cancer --- p.14 / Chapter 2.4 --- Prevention and Treatment --- p.16 / Chapter 2.5 --- Molecular alterations in endometrial cancer --- p.17 / Chapter 2.5.1 --- Genetic alterations in endometrial cancer --- p.18 / Chapter 2.5.1.1 --- Oncogene activation --- p.19 / Chapter 2.5.1.2 --- Tumor suppressor gene inactivation --- p.20 / Chapter 2.5.1.2.1 --- Mutation and loss of heterozygosity of tumor suppressor genes in endometrial cancer --- p.22 / Chapter 2.5.2 --- Epigenetic alterations --- p.27 / Chapter 2.5.2.1 --- CpG islands methylation --- p.29 / Chapter 2.5.2.2 --- de novo methylation --- p.29 / Chapter 2.5.2.3 --- Detection of gene promoter hypermethylation --- p.31 / Chapter 2.5.2.4 --- Epigenetic alteration in endometrial cancer --- p.31 / Chapter 2.5.2.5 --- Promoter hypermethylation of tumor suppressor genes in other cancers --- p.39 / Chapter 2.5.3 --- Microsatellite instability --- p.42 / Chapter Chapter 3 --- The objectives of study --- p.45 / Chapter Chapter 4 --- Materials and Methods --- p.46 / Chapter 4.1 --- Samples --- p.46 / Chapter 4.1.1 --- Formalin fixed paraffin embedded tissues --- p.46 / Chapter 4.1.2 --- Cell lines --- p.46 / Chapter 4.2 --- Histological grading and staging of samples --- p.47 / Chapter 4.3 --- Microdissection on tissue sections --- p.47 / Chapter 4.4 --- Extraction of nucleic acid / Chapter 4.4.1 --- Extraction of DNA from paraffin-embedded tissues --- p.48 / Chapter 4.4.2 --- Extraction of DNA from cell lines --- p.49 / Chapter 4.5 --- DNA methylation analysis --- p.49 / Chapter 4.5.1 --- Overview of Methylation-Specific PCR (MSP) --- p.49 / Chapter 4.5.2 --- Bisulfite modification of DNA --- p.50 / Chapter 4.5.3 --- Methylation specific PCR (MSP) --- p.51 / Chapter 4.6 --- Microsatellite Analysis --- p.53 / Chapter 4.7 --- Statistical analysis --- p.56 / Chapter Chapter 5 --- Results / Chapter 5.1 --- Clinical-pathological features of endometrioid adenocarcinoma --- p.57 / Chapter 5.2 --- Promoter hypermethylation in endometrial cancer --- p.57 / Chapter 5.3 --- Microsatellite status (MSI) analysis --- p.65 / Chapter Chapter 6 --- Discussion / Chapter 6.1 --- Promoter hypermethylation in endometrial cancer --- p.71 / Chapter 6.1.1 --- Concurrent hypermethylation of multiple genesin endometrioid adenocarcinoma and its precursor lesions --- p.72 / Chapter 6.1.1.1 --- Promoter hypermethylation of E-cad --- p.73 / Chapter 6.1.1.2 --- Promoter hypermethylation of APC --- p.73 / Chapter 6.1.1.3 --- Promoter hypermethylation of MGMT --- p.74 / Chapter 6.1.1.4 --- Promoter hypermethylation of RASSF1A --- p.75 / Chapter 6.1.1.5 --- Promoter hypermethylation of hMLH-1 --- p.76 / Chapter 6.1.1.6 --- Promoter hypermethylation in ECA coexisting with hyperplasia and not coexisting with hyperplasia --- p.77 / Chapter 6.1.2 --- Promoter hypermethylation in SCA --- p.77 / Chapter 6.2 --- Microsatellite status analysis --- p.78 / Chapter 6.2.1 --- MSI in endometrial cancer --- p.78 / Chapter 6.2.2 --- MSI and concurrent promoter hypermethylation --- p.79 / Chapter 6.2.3 --- MSI and promoter hypermethylation of hMLH-1 --- p.80 / Chapter Chapter 7 --- Conclusion --- p.81 / Further studies --- p.82 / References --- p.83

子宮内膜癌是最常见的妇科恶性肿瘤，其中有80%属于子宮内膜腺样癌，这一癌症发病的分子机制尚未清楚。研究表明，癌基因表达异常或者功能异常在肿瘤的发生发展过程中具有重要的作用。另一方面，抑癌基因在肿瘤细胞中特异性甲基化失活通常导致细胞恶性转变和肿瘤的发生。本实验将阐明癌基因阴阳1和抑癌基因PCDH10在人类子宮内膜腺样癌发病中的作用。 / 本实验第一部分研究多功能转录因子阴阳1（YY1）在子宮内膜腺样癌发病过程中的作用。首先本实验证实YY1在子宮内膜腺样癌临床标本和癌细胞系中均明显表达上调，并且上调的程度与肿瘤的FIGO分期相关。接着体外细胞培养和裸鼠荷瘤模型的实验均提示抑制YY1 表达可抑制癌细胞增殖和体外迁移，而过表达YY1则促进癌细胞增殖。这些结果表明YY1在子宮内膜腺样癌发病中具有促进作用。进一步全细胞基因组转录谱分析提示YY1 介入子宮内膜腺样发病的各个方面，并通过抑制抑癌基因APC的表达发挥发挥重要作用。深入的分子机制研究发现一个新的表观抑制作用模型：YY1可募集EZH2等多梳蛋白到APC启动子区并导致后者组蛋白3赖氨酸27上三甲基化，从而抑制APC基因转录。此外，本实验还发现YY1在子宮内膜腺样癌的表达增高是由于微小RNA，miR-193a-5p，在此癌中表达下降所导致的。所以，本实验第一部分的结果揭示了miR-193a-5p-YY1-APC这条全新的信号通路在子宮内膜腺样癌发病中发挥重要作用，并可作为潜在的治疗靶点。 / 本实验第二部分鉴定出PCDH10作为子宮内膜腺样癌一个新的抑癌基因。通过5-氮杂-2'-氧胞嘧啶处理和亚硫酸氢钠测序的方法，我们证实抑癌基因PCDH10在子宮内膜腺样癌中失活是由于其启动子区DNA甲基化所致，并且这种DNA甲基化介导的PCDH10表达沉默在子宮内膜腺样癌临床标本和癌细胞系中很常见，但不存在于正常子宮内膜组织。另外，在子宮内膜腺样癌细胞系体外实验中恢复PCDH10的表达可抑制细胞增殖、单细胞克隆形成，促进细胞凋亡。 同时在体实验荷瘤模型中恢复PCDH10的表达也可抑制肿瘤细胞增殖，这些结果与其肿瘤抑制功能相符。 / 总之，本实验结果阐明了YY1和PCDH10在子宮内膜腺样癌发病过程中新的作用，拓展了子宮内膜腺样癌发病分子机制的研究并为其药物治疗提供了潜在的靶点。 / Endometrial cancer is the most common gynecologic malignancy and about 80% of these cancers are endometrial Endometrioid carcinoma (EEC). The molecular mechanisms underlying EEC tumorigenesis are under-explored. Aberrant expression and function of oncogenes promote tumor development by modulating many aspects of tumor cell growth. On the other hand, tumor specific promoter methylation on tumor suppressor genes (TSG), which are generally unmethylated in normal cells, usually initiate and promote malignant transformation and cancer initiation. Our study aims to characterize the functions of an oncogenic transcription factor Yin Yang 1 (YY1) and a novel tumor suppressor gene PCDH10 in Human Endometrioid Endometrial Adenocarcinoma. / In the first part of our study, we investigated the function of a multifunctional TF, YY1 in EEC tumorigenesis. We demonstrated YY1 is up-regulated in EEC cell lines and primary tumors and its expression is associated with FIGO stages. Depletion of YY1 inhibits EEC cell proliferation and migration both in vitro and in vivo whereas over-expression of YY1 promotes EEC cell growth. These results suggest that YY1 functions as an onocogenic factor in EEC. Transcriptome analysis revealed a significant effect of YY1 on critical aspects of EEC tumorigenesis and its down-regulation of APC transcripts. Further mechanistic investigation uncovered a new epigenetic silencing mode of Adenomatosis Polyposis Coli (APC) by YY1 through recruitment of EZH2 and trimethylation of histone 3 lysine 27 in its promoter region. Additionally, YY1 over-expression was found to be a consequence of miR-193a-5p down-regulation through direct miR-193a-5p-YY1 interplay. Our results therefore established a novel miR-193a-5p-YY1-APC regulatory axis contributing to EEC development, which may serve as future intervention target. / In the second part of our study, we identified PCDH10 as a novel tumor suppressor gene in EEC. By using bisulfate genomic sequencing combined with pharmacologic demethylation drug treatment, we elucidated that PCDH10 inactivation in EEC is a consequence of DNA hypermethylation on its promoter region. Further study suggested that hypermethylation-mediated PCDH10 silencing was a common event in EEC cell lines and clinical samples, but not in normal endometrial tissues. Restoration of PCDH10 expression in EEC cells suppressed cell proliferation, inhibited single cell colony formation and induced cell apoptosis; moreover, overexpression of PCDH10 inhibited EEC xenograft tumor growth in vivo.These results suggest PCDH10 acts as a tumor suppressor. / Together, our results reveal the novel functions of YY1 and PCDH10 in EEC. These findings add novel insights into the molecular mechanisms of EEC development and progression, which may serve as potential therapeutic targets for this disease. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Yang, Yihua. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 170-186). / Abstracts also in Chinese. / TITLE --- p.I / ABSTRACT --- p.III / ACKNOWLEDGEMENTS --- p.VII / PUBLICATION --- p.IX / ABBREVIATIONS --- p.X / LIST OF FIGURES --- p.XIII / LIST OF TABLES --- p.XVI / TABLES OF CONTENT --- p.XVII / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Endometrioid Endometrial Adenocarcinoma (EEC) --- p.1 / Chapter 1.1.1 --- Epidemiology --- p.2 / Chapter 1.1.2 --- Etiology and risk factors --- p.3 / Chapter 1.1.3 --- Treatment and prognosis --- p.8 / Chapter 1.1.4 --- Molecular Mechanisms --- p.9 / Chapter 1.1.5 --- APC and Wnt/β-catenin signaling pathway --- p.12 / Chapter 1.1.6 --- Summary --- p.18 / Chapter 1.2 --- Epigenetic modifications and EEC --- p.20 / Chapter 1.2.1 --- Epigenetic modifications --- p.20 / Chapter 1.2.2 --- Epigenetic and cancer --- p.21 / Chapter 1.2.5 --- Summary --- p.30 / Chapter Chapter 2 --- Material and Method --- p.31 / Chapter 2.1 --- Tissue samples --- p.31 / Chapter 2.2 --- Cell culture --- p.32 / Chapter 2.3 --- Cell proliferation assays --- p.33 / Chapter 2.4 --- Cell migration assay --- p.34 / Chapter 2.5 --- 3-deazaneplanocin A (Dznep) or 5-aza-2'-deoxycytidine (5-aza) treatment --- p.34 / Chapter 2.6 --- Computational prediction --- p.35 / Chapter 2.7 --- Cell cycle assay --- p.35 / Chapter 2.8 --- Apoptosis assay --- p.36 / Chapter 2.9 --- Total RNAs, Total proteins and Genomic DNA extraction --- p.36 / Chapter 2.10 --- Bisulfite Genomic Sequencing --- p.38 / Chapter 2.11 --- Oligonucleotides --- p.39 / Chapter 2.12 --- RT-PCR, Semi-quantitative PCR and Real-time PCR --- p.41 / Chapter 2.13 --- microRNA validation --- p.43 / Chapter 2.14 --- Plasmid construction --- p.43 / Chapter 2.15 --- Transfection --- p.45 / Chapter 2.16 --- Luciferase reporter assay --- p.45 / Chapter 2.17 --- Western blotting --- p.46 / Chapter 2.18 --- Immunofluorescence ( IF ) --- p.48 / Chapter 2.19 --- Immunohistochemistry (IHC) --- p.50 / Chapter 2.20 --- ChIP assay --- p.53 / Chapter 2.21 --- Sequencing and base calling --- p.55 / Chapter 2.22 --- Read mapping to genome with splice-aware aligner sequenced --- p.55 / Chapter 2.23 --- Xenograft mouse model --- p.55 / Chapter 2.24 --- Statistical analysis --- p.57 / Chapter Chapter 3 --- Yin Yang 1 Plays an Oncogenic Role in Human Endometrioid Endometrial Adenocarcinoma --- p.58 / Chapter 3.1 --- YIN YANG 1(YY1) --- p.58 / Chapter 3.1.1 --- YY1 structure --- p.58 / Chapter 3.1.2 --- YY1 function --- p.59 / Chapter 3.1.3 --- YY1 and epigenetic --- p.61 / Chapter 3.1.4 --- YY1 and cancer --- p.62 / Chapter 3.1.5 --- Regulation of YY1 expression and activity --- p.66 / Chapter 3.2 --- Results --- p.68 / Chapter 3.2.1 --- YY1 is up-regulated in EEC lines and localizes in nuclei of EEC cells --- p.68 / Chapter 3.2.2 --- YY1 expression level is associated with EEC clinicopathological features --- p.72 / Chapter 3.2.3 --- Knock-down of YY1 by RNAi inhibits EEC cell proliferation and migration --- p.77 / Chapter 3.2.4 --- Ectopic expression of YY1 promotes EEC cell proliferation --- p.84 / Chapter 3.2.5 --- YY1 does not affect EEC cell cycle and cell apoptosis --- p.91 / Chapter 3.2.6 --- Genome-wide characterization of YY1-mediated transcriptome changes --- p.94 / Chapter 3.2.7 --- Gene Ontology analysis of YY1 targets on EEC tumorigenesis --- p.98 / Chapter 3.2.8 --- YY1 inhibits APC gene expression and functions --- p.101 / Chapter 3.2.9 --- YY1 inhibits APC expression through recruiting EZH2 and causing H3K27me3. --- p.105 / Chapter 3.2.10 --- Knock-down of YY1 does not change DNA methylation status of CpG island of APC gene --- p.117 / Chapter 3.2.11 --- SiYY1 oligo injection inhibits tumor grows in vivo --- p.119 / Chapter 3.2.12 --- miR-193a-5p is down-regulated in EEC cell lines and clinical samples --- p.126 / Chapter 3.2.13 --- miR-193a-5p targets YY1 3’UTR and inhibits YY1 expression --- p.128 / Chapter 3.2.14 --- miR-193a-5p inhibits tumor grow in vivo --- p.133 / Chapter 3.3 --- Discussion --- p.136 / Chapter 3.3.1 --- YY1 oncogenic functions in EEC --- p.136 / Chapter 3.3.2 --- YY1 epigenetically silences APC --- p.138 / Chapter 3.3.3 --- miR-193a-5p down-regulates YY1 in EEC --- p.139 / Chapter 3.4 --- Conclusion --- p.141 / Chapter Chapter 4 --- The tumor suppressive functions of PCDH10 in Human Endometrioid Endometrial Adenocarcinoma --- p.143 / Chapter 4.1 --- Introduction --- p.143 / Chapter 4.1.1 --- PCDH10 structure and function --- p.143 / Chapter 4.1.2 --- PCDH10 and tumor --- p.146 / Chapter 4.1 --- Results --- p.149 / Chapter 4.2.1 --- PCDH10 is down-regulated in EEC cell lines and clinical samples --- p.149 / Chapter 4.2.2 --- PCDH10 is hypermethylated in EEC cell lines and clinical samples --- p.150 / Chapter 4.2.3 --- Pharmacologic demethylation restores PCDH10 expression in EEC cell lines --- p.152 / Chapter 4.2.4 --- Ectopic over-expression of PCDH10 inhibits EEC cell proliferation --- p.154 / Chapter 4.2.5 --- PCDH10 over-expression induces EEC cell apoptosis --- p.161 / Chapter 4.2.6 --- PCDH10 over-expression inhibits tumor grows in vivo --- p.166 / Chapter 4.3 --- Discussion and future plan --- p.169 / REFERENCE --- p.170

The endometrium is receptive to the embryo during a restricted window in the mid-secretory phase. My objectives were to develop a minimally-invasive endometrial sampling method for gene expression profiling, and to identify genes differentially expressed in the receptive phase. Twenty-three normo-ovulatory women underwent uterine fluid aspiration during the pre-receptive (LH+2) and receptive (LH+7) phase of the same natural cycle. RNA was extracted, reverse transcribed, amplified and hybridized to whole-genome microarrays. Unsupervised hierarchical clustering revealed self-segregation of pre-receptive and receptive samples. Importantly, profiling by uterine fluid aspiration was representative of biopsy. An unpaired t-test with a false discovery rate of 0.05 and a Δ threshold of 4-fold identified 245 unique transcripts as differentially expressed in the receptive phase. NanoString analysis validated 96% of these genes. This approach will now allow us to correlate expression of these candidate biomarkers to implantation outcomes, towards the development of clinical assays predictive for endometrial receptivity.

The endometrium is receptive to the embryo during a restricted window in the mid-secretory phase. My objectives were to develop a minimally-invasive endometrial sampling method for gene expression profiling, and to identify genes differentially expressed in the receptive phase. Twenty-three normo-ovulatory women underwent uterine fluid aspiration during the pre-receptive (LH+2) and receptive (LH+7) phase of the same natural cycle. RNA was extracted, reverse transcribed, amplified and hybridized to whole-genome microarrays. Unsupervised hierarchical clustering revealed self-segregation of pre-receptive and receptive samples. Importantly, profiling by uterine fluid aspiration was representative of biopsy. An unpaired t-test with a false discovery rate of 0.05 and a Δ threshold of 4-fold identified 245 unique transcripts as differentially expressed in the receptive phase. NanoString analysis validated 96% of these genes. This approach will now allow us to correlate expression of these candidate biomarkers to implantation outcomes, towards the development of clinical assays predictive for endometrial receptivity.

Endometrial cancer is one of the most significant gynaecological malignancies that affect women from New Zealand and the rest of the world. One of the critical stages in the development of a tumour is the onset of hypoxia. The malignancy responds by having raised levels of Hypoxia Inducible Factor (HIF) that in turn induces increased production of Vascular Endothelial Growth Factor (VEGF). VEGF is a potent angiogenic factor that will mediate vascular supply of nutrients and oxygen to the developing tumour. The aim of this study was to investigate whether two compounds found in extracts of plant materials, Resveratrol (Resveratrol) and Epigallocatechin gallate (EGCG), altered the levels of VEGF in the supernatant of cultured endometrial cancer cells. Resveratrol is a phytoalexin that is found in many foods, such as grapes, nuts and berries, as well as in high concentrations in some red wines. 100 µM of resveratrol was added to cell cultures for 24 hours. VEGF levels in the supernatant were then analysed using ELISA. Resveratrol was found to have significant inhibitory effects in both primary endometrial cancer cell cultures and immortalised endometrial cancer cell cultures. Resveratrol was also shown to reverse the increase in VEGF caused by the hypoxia mimic cobalt chloride (CoCl₂). Epigallocatechin gallate (EGCG) is an antioxidant catechin extracted from green tea. The effect of EGCG was analysed using the same method as for resveratrol. 100 µM of EGCG was also shown to have a significant inhibitory effect on the level of VEGF in the supernatant of cultured endometrial cancer cells, as well as reducing the effect of CoCl₂. These results suggest that selected food compounds, resveratrol and EGCG, can reduce VEGF levels by inhibiting HIF. Further investigation This may have anti-tumour effects in women with endometrial cancer.

Diss. (sammanfattning) Umeå : Univ., 2003. / Härtill 4 uppsatser.