It is currently poorly understood how hormones like oestrogen can elicit a rapid non-genomic response from cells via their traditional nuclear receptors. It has been known for some time that some cells, although lacking the canonical oestrogen receptors (ERs), still display rapid non-genomic signalling in response to oestrogen. Recently, a G protein coupled receptor; GPR30 (or GPER) and an alternative splice variant of the traditional ERα (ERα36) have been proposed as alternative oestrogen receptors that may mediate these non-genomic effects. This study investigated the presence and location of GPR30 and ERα36 in breast and endometrial cancer cells. We also investigated their possible roles in cancer cell migration. Using a combination of cell migration assays, gene silencing siRNAs, actin polymerisation measurements and immuno-fluorescence we have demonstrated that SKBr3 cells (a ER receptor negative, breast cancer cell line) migrate towards the GPR30/ERα36 agonist G-1; an effect that is attenuated by G-15, a GPR30/ERα36 antagonist and pERK inhibitors, but not by ROCK inhibitors. We have also demonstrated that G-1 activates a rapid signalling pathway involving changes in the actin cytoskeleton. A similar migration response is seen in endometrial cancer cells. However, we have also discovered that another breast cancer, ER receptor negative cell line, MDA-MB-231, although expressing both GPR30 and ERα36 does not display this migratory behaviour. We propose and provide evidence that the migratory signalling pathway via GPR30 and ERα36 involves the activation of the HER 2 receptor which the MDA-MB-231 cell line lacks. These findings may progress the development of new therapeutics targeting ER negative breast cancer tumors.

published_or_final_version / Medicine / Master / Doctor of Medicine

Establishment of pregnancy in ruminants requires blastocyst development to form an elongated filamentous conceptus that produces interferon tau (IFNT), the pregnancy recognition signal, and initiate implantation. Blastocyst growth and development is dependent upon secretions from the uterine endometrium. An early increase in post-ovulatory circulating levels of progesterone (P4) stimulates blastocyst growth and conceptus elongation in ruminants. Microarray analysis was used to identify candidate P4-regulated genes and regulatory networks in the endometrium that govern peri-implantation blastocyst/conceptus growth and development. The first study was conducted to validate effects of P4 and/or pregnancy on expression of candidate genes identified by microarray analysis. The genes included: ANGPTL3, CHGA, CLEC4E, CXCL14, EFNA1, EFNB1, FABP3, IFNG, IL6, LGALS3, PTH, RBP4, SLIT2, SLIT3, and VWF. Early P4 treatment up-regulated CXCL14 gene expression in Day 9 ovine endometrium compared to control endometrium, and FABP3, IFNG, IL6 and LGALS3 in Day 12 early P4-treated ovine endometrium. Expression of ANGPTL3, CHGA, CXCL14, EFNA1, EFNB1, LGALS3 and RBP4 was affected by day of pregnancy. Treatment of ewes with P4+RU486 (P4 receptor antagonist) reduced expression of ANGPTL3, CHGA, EFNA1, EFNB1, FABP3, IFNG, IL6, LGALS3, RBP4, and SLIT2, SLIT3 and VWF in comparison to Day 12 P4-treated endometrium. The second study evaluated expression of genes identified by microarray analysis in endometrium from pregnant and cyclic ewes. Genes evaluated included those from the first study. ANGPTL3, CHGA, CXCL14, EFNA1, EFNB1, IFNG, LGALS3, PTH, RBP4, SLIT2, SLIT3 and VWF were affected by day, status and/or their interaction between Days 10 and 16. Of note, FABP3 increased 21-fold between Days 14 to 18 of pregnancy, and IL6 increased 37-fold between Days 14 to 20 of pregnancy. In situ hybridization analysis detected FABP3 mRNA in both luminal and superficial glandular epithelia of pregnant ewes and trophectoderm, whereas IL6 mRNA was detected in immune cells within uterine luminal epithelium and glandular epithelium and trophectoderm. Collectively, these results identify candidate genes encoding for biologically active molecules that regulate growth and development of the ovine conceptus during the peri-implantation period of pregnancy.

Thesis (Ph. D.)--University of Hong Kong, 2008. / Also available in print.

Proefschrift Universiteit van Amsterdam.

Thesis (doctoral)--Utrecht, 1984.

Thesis (doctoral)--München, 1979.

Proefschrift Universiteit Maastricht. / Auteursnaam op omslag: Hanny Pijnenborg. Met lit. opg. - Met samenvatting in het Nederlands.

Proefschrift Universiteit Maastricht. / Met bibliogr., lit. opg. - Met samenvatting in het Nederlands en Turks.

Cell adhesion proteins osteopontin, CD44 and integrin alphaVbeta3 interact to form an adhesion complex between the embryo and endometrial surface forming an attachment that can lead to implantation. Whilst receptivity has been investigated extensively, the expression of this adhesion complex has yet to be studied simultaneously in the endometrium. This thesis establishes the expression of the adhesion complex in fertile and infertile endometrium. In addition the regulation of the adhesion complex components by distinct signalling pathways and the key regulators estrogen receptor, nuclear factor kappa B and signal transducer and activator of transcription 1 have been investigated in endometrial cell lines. Objectives: To establish the expression profile of adhesion complex components in samples obtained from fertile and infertile women. To model in vitro hormonal regulation of adhesion complex components to mimic estrogen and progesterone stimulus in the menstrual cycle. To determine if adverse environments common to poly cystic ovarian syndrome and endometriosis affect uterine expression of the adhesion complex via high glucose and pro-inflammatory cytokines. To investigate the direct regulation of Osteopontin and CD44 by estrogen and cytokine signalling through estrogen receptor ?, nuclear factor kappa B and signal transducer and activator of transcription 1. Methodology: Investigation of human biopsies and cell line models by immunohistochemistry, quantitative polymerase chain reaction, chromatin immunoprecipitation and enzyme linked immunosorbent assay. Conclusions: Adverse uterine environments including high glucose and pro-inflammatory cytokines may regulate the expression of the adhesion complex, and contribute to a lack of endometrial receptivity in endometriosis and poly cystic ovarian patients. CD44, ITGAV and ITGB3 levels may be used as markers for loss of receptivity in unexplained infertility.