Significance of progesterone receptors (PR-A and PR-B) expression as predictors for relapse after successful therapy of endometrial hyperplasia: a retrospective cohort study

This is the peer reviewed version of the following article: Sletten, E.T., Arnes, M., Lyså, L.M., Larsen, M. & Ørbo, A. (2019). Significance of progesterone receptors (PR-A and PR-B) expression as predictors for relapse after successful therapy of endometrial hyperplasia: a retrospective cohort study. BJOG: an International Journal of Obstetrics and Gynaecology, 126(7), 936-943, which has been published in final form at https://doi.org/10.1111/1471-0528.15579. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.Objective - After successful progestin therapy for endometrial hyperplasia (EH), the risk of relapse remains. We aimed to assess if immunohistochemical (IHC) expression of progesterone receptor isoforms, PR‐A and PR‐B, in endometrial glands and stroma in pre‐treatment endometrial biopsies was related to relapse of EH. Design and setting - Biopsy material originated from women with low‐risk and medium‐risk EH recruited to a recent Norwegian multicentre randomised trial. Participants (n = 153) had been treated for 6 months with three different progestin regimens. Population - One hundred and thirty‐five of the 153 women achieved therapy response and underwent follow up for 24 months after therapy withdrawal. Fifty‐five women relapsed during follow up. Pre‐treatment endometrial biopsies from 94 of the 135 responding women were available for IHC staining. Methods - Immunohistochemical staining was performed separately for PR‐A and PR‐B and IHC expression was evaluated in endometrial glands and stroma by a histological score (H‐score) using light microscopy. Main outcome measure - Immunohistochemical expression of PR‐A and PR‐B in endometrial glands and stroma in women with or without relapse of EH. Results - Low PR‐A in endometrial glands (P = 0.013) and stroma (P 1 (19%; P < 0.001). Conclusion - Immunohistochemical expression of PR‐A and PR‐B in pre‐treatment endometrial biopsy proves valuable as a predictor of relapse in EH

Downregulation of TFPI in breast cancer cells induces tyrosine phosphorylation signaling and increases metastatic growth by stimulating cell motility

<p>Abstract</p> <p>Background</p> <p>Increased hemostatic activity is common in many cancer types and often causes additional complications and even death. Circumstantial evidence suggests that tissue factor pathway inhibitor-1 (TFPI) plays a role in cancer development. We recently reported that downregulation of TFPI inhibited apoptosis in a breast cancer cell line. In this study, we investigated the effects of TFPI on self-sustained growth and motility of these cells, and of another invasive breast cancer cell type (MDA-MB-231).</p> <p>Methods</p> <p>Stable cell lines with TFPI (both α and β) and only TFPIβ downregulated were created using RNA interference technology. We investigated the ability of the transduced cells to grow, when seeded at low densities, and to form colonies, along with metastatic characteristics such as adhesion, migration and invasion.</p> <p>Results</p> <p>Downregulation of TFPI was associated with increased self-sustained cell growth. An increase in cell attachment and spreading was observed to collagen type I, together with elevated levels of integrin α2. Downregulation of TFPI also stimulated migration and invasion of cells, and elevated MMP activity was involved in the increased invasion observed. Surprisingly, equivalent results were observed when TFPIβ was downregulated, revealing a novel function of this isoform in cancer metastasis.</p> <p>Conclusions</p> <p>Our results suggest an anti-metastatic effect of TFPI and may provide a novel therapeutic approach in cancer.</p

Knowledge of the Climate or Action Competence for Sustainable Development? A Case Study of How Students Are Taught at Mímisbrunnr Klimapark 2469

I ei verd med stadige utfordringar, særleg med omsyn til klima og miljø, treng vi engasjerte verdsborgarar som har dei reiskapane dei treng for å løyse utfordringane. Da kjem ein fort til spørsmål om kva desse reiskapane er – kva skal til for å fremja berekraftig utvikling? Eg har i denne mastergradsoppgåva sett på korleis Mímisbrunnr Klimapark 2469 legg opp si undervisning, for å finne svar på problemstillinga mi: Kunnskap om klima eller handlingskompetanse for berekraftig utvikling – kva legg Mímisbrunnr Klimapark 2469 vekt på, og kva lærer elevane? Teorien eg har brukt, er litterataur om utdanning for berekraftig utvikling (UBU), utvalde delar av planverket i norsk skule, teori om kompetansar- og kompetanseomgrepet, uteundervisning og undervisning på museum, samt teori om menneske i møte med klimabodskapen. UBU er eit omdiskutert fagområde, og utdrag av kritikken mot det er òg med i teorien. For å finne svar på det eg lurte på har eg brukt fire ulike metodar: intervju med ein tilsett, observasjon av ei skuleklasse på vitjing, fokusgruppeintervju med elevar frå klassa eg observerte og undersøkingar av Klimaparken sine interne dokument. Analysen vart gjennomført ved bruk av ein analysemodell som viser i kor stor grad undervisningsopplegg fremjar berekraft. Resultata viste at Klimaparken i dokumenta sine har intensjonar om eit undervisningsopplegg som fremjar berekraftig utvikling med kunnskap om, handlingar for, erfaringar i og erfaringar som berekraftig utvikling, utan at observasjonen tydeleg viser at elevane får utvikla si handlingskompetanse, eller andre kompetansar for UBU. Det som elevane derimot har etter undervisningsopplegget, er kunnskap. Dette er kunnskap som held eit høgt fagleg nivå om mange ulike tema, men elevane klarer ikkje tydeleg å sjå samanhengane mellom temaa. Eit undervisningsopplegg i UBU må leggje vekt på både undervisning for, i, om og som berekraftig utvikling. Undersøkingane viser at undervisningsopplegget og målet som Klimaparken har om å fremja berekraftig utvikling, har svakheiter. Med støtte i teori, har eg skissert korleis svakheitene kan reduserast for at opplegget i større grad kan fremja UBU.In a world full of challenges, especially in the natural environment, committed citizens who possesses to take action and solve problems are desirable. This forms the basis of the question that my thesis will attempt to answer: What are the tools needed for achieving suitable development? To find an answer to my thesis statement: knowledge of the climate or action competence for sustainable development – What is the main focus of Mímisbrunnr Klimapark 2469, and what do the students learn there? – I have researched how Mímisbrunnr Climate Park 2469’s teaching is planned and carried out. The theory used in this thesis is based on education for sustainable development (ESD), chosen parts of the curriculum in Norwegian school, theory on competences and the competence term, teaching outdoors and in museums, in addition to theory about humans psychological reactions to the climate challenges. EDS is a debated term and field and that is why the theory of this thesis also includes some of the critics to the field. In order to answer my thesis statement I have used four different research methods: interview, observation, focus group interview and research of documents at the Climate Park. The analysis was done by using an analysis model to search for ESD in the teaching plan. The results show that the documents of the Climate Park had intentions for ESD by focusing on knowledge about, actions for, practice in and experiences as sustainable development. The observation shows few signs of focus on the students actions or their action competence. In the interview the students are talking about knowledge that they have acquired. They have acquired certain subjects, but there are signs that they are missing out on the connections between the subjects. This might imply that the teaching plan and the goals, which the Climate Park has in-place for ESD has some weaknesses. A teaching plan should contain knowledge about, actions for, practice in and experiences as sustainable development to be fully in line with ESD. The findings suggest that the teaching plan can be adjusted to more successfully providing ESD. With support from the theory presented I give a short example of how this can be done.M-LU

Intrauterine progestin therapy as a new approach to premalignant endometrial polyps: A prospective observational study

Background/Aim: Endometrial hyperplastic polyps (EHP) may progress to endometrial carcinoma (EC) if left untreated. We aimed to prospectively investigate the efficacy of the low-dose levonorgestrel intrauterine system (LNG-IUS) as therapy for EHP with malignant potential. Patients and Methods: In total, 37 women with EHP underwent therapy with LNG-IUS containing 13.5 mg levonorgestrel for six months or 4-10 weeks depending on whether the EHP was characterized (by D-score analysis) as low- to medium-risk (n=33) or high-risk (n=4) of coexistent or future EC. Therapy response was defined as complete clearance of hyperplastic glands in post-therapy endometrial biopsy. Results: All women with low- to medium-risk EHP obtained therapy response, whereas only 1 out of 4 with high-risk EHP responded to therapy. None of the women were diagnosed with EC during the study and no serious adverse events occurred. Conclusion: Low-dose LNG-IUS represents a promising therapy for selected women with EHP

TFPIα and TFPIβ are expressed at the surface of breast cancer cells and inhibit TF-FVIIa activity

<p>Abstract</p> <p>Background</p> <p>Tissue factor (TF) pathway inhibitor-1 (TFPI) is expressed in several malignant tissues- and cell lines and we recently reported that it possesses anti-tumor effects in breast cancer cells, indicating a biological role of TFPI in cancer. The two main splice variants of TFPI; TFPIα and TFPIβ, are both able to inhibit TF-factor VIIa (FVIIa) activity in normal cells, but only TFPIα circulates in plasma. The functional importance of TFPIβ is therefore largely unknown, especially in cancer cells. We aimed to characterize the expression and function of TFPIα, TFPIβ, and TF in a panel of tumor derived breast cancer cell lines in comparison to normal endothelial cells.</p> <p>Methods</p> <p>TFPIα, TFPIβ, and TF mRNA and protein measurements were conducted using qRT-PCR and ELISA, respectively. Cell-associated TFPI was detected after phosphatidylinositol-phospholipase C (PI-PLC) and heparin treatment by flow cytometry, immunofluorescence, and Western blotting. The potential anticoagulant activity of cell surface TFPI was determined in a factor Xa activity assay.</p> <p>Results</p> <p>The expression of both isoforms of TFPI varied considerably among the breast cancer cell lines tested, from no expression in Sum149 cells to levels above or in the same range as normal endothelial cells in Sum102 and MDA-MB-231 cells. PI-PLC treatment released both TFPIα and TFPIβ from the breast cancer cell membrane and increased TF activity on the cell surface, showing TF-FVIIa inhibitory activity of the glycosylphosphatidylinositol- (GPI-) anchored TFPI. Heparin treatment released TFPIα without decreasing the cell surface levels, thus indicating the presence of intracellular storage pools of TFPIα in the breast cancer cells.</p> <p>Conclusion</p> <p>GPI-attached TFPI located at the surface of breast cancer cells inhibited TF activity and could possibly reduce TF signaling and breast cancer cell growth locally, indicating a therapeutic potential of the TFPIβ isoform.</p

Global gene expression analysis reveals a link between NDRG1 and vesicle transport

-N-myc downstream-regulated gene 1 (NDRG1) is induced by cellular stress such as hypoxia and DNA damage, and in humans, germ line mutations cause Charcot-Marie-Tooth disease. However, the cellular roles of NDRG1 are not fully understood. Previously, NDRG1 was shown to mediate doxorubicin resistance under hypoxia, suggesting a role for NDRG1 in cell survival under these conditions. We found decreased apoptosis in doxorubicin-treated cells expressing NDRG1 shRNAs under normoxia, demonstrating a requirement for NDRG1 in apoptosis in breast epithelial cells under normal oxygen pressure. Also, different cellular stress regimens, such as hypoxia and doxorubicin treatment, induced NDRG1 through different stress signalling pathways. We further compared expression profiles in human breast epithelial cells ectopically over-expressing NDRG1 with cells expressing NDRG1 shRNAs in order to identify biological pathways where NDRG1 is involved. The results suggest that NDRG1 may have roles connected to vesicle transport

Indirect regulation of TFPI-2 expression by miR-494 in breast cancer cells

Abstract TFPI-2 has been shown to be involved in breast cancer pathogenesis by inhibiting extracellular matrix degradation, and low levels are associated with disease progression. As microRNA-494 (miR-494) protects against breast cancer progression, we investigated whether miR-494 is involved in the regulation of TFPI-2 in MCF-7 breast cancer cells. TFPI-2 mRNA and protein levels increased after transfection with miR-494 mimic, and TFPI-2 mRNA and miR-494 levels correlated positively in tumors from breast cancer patients. No specific binding sites for miR-494 in the 3′-untranslated region (UTR) of TFPI2 were identified; however, miR-494 was predicted in silico to bind 3′-UTR of the transcription factors AHR and ELF-1, which have potential binding sites in the TFPI2 promoter. ELF-1 mRNA was downregulated whereas AHR mRNA levels were upregulated after transfection with miR-494 mimic. Knockdown of ELF-1 and AHR increased and reduced TFPI-2 mRNA levels, respectively. Increased luciferase activity was seen when TFPI-2 promoter constructs containing the potential AHR or ELF-1 binding sites were co-transfected with miR-494 mimic. In conclusion, TFPI-2 mRNA levels were upregulated by miR-494 in MCF-7 breast cancer cells most likely by an indirect association where miR-494 targeted the transcription factors AHR and ELF-1. This association was supported in a breast cancer cohort