Treatment Strategies For Endometrial Cancer Research Articles

Locked nucleic acid-inhibitor of miR-205 decreases endometrial cancer cells proliferationin vitroandin vivo

Pathogenesis of endometrial cancer has been connected with alterations of microRNA expression and in particular miR-205 up–regulation was consistently reported in this carcinoma. Presented study aimed to investigate if inhibition of miR-205 expression using LNA-modified-nucleotide would attenuate endometrial cancer cells proliferation in vitro and in vivo.In the course of the study we found that the proliferation of endometrial cancer cells (HEC-1-B, RL-95, KLE, Ishikawa) transfected with LNA-miR-205-inhibitor and evaluated using real time cell monitoring as well as standard cell proliferation assay, was significantly decreased. Next, LNA-miR-205-inhibitor was used to assess the in vivo effects of miR-205 inhibition of endometrial cancer growth. Cby.Cg-Foxn1<nu>/cmdb mice bearing endometrial cancer xenografts were intraperitoneally injected with nine dosages of 25mg/kg of miR-205-LNA-inhibitor or scramble control or phosphatase buffered saline and were observed for 32 days. We found that systemic administration of miR-205-LNA-inhibitor was technically possible, and exerted inhibitory effect on endometrial cancer xenograft growth in vivo with only mild toxic effects in treated animals.In conclusion our results suggest that systemic delivery of miR-205-LNA-inhibitor is feasible, devoid of significant toxicity, and could be a promising treatment strategy for endometrial cancer. Therefore it warrants further studies in other animal models.

Abstract 2994: Orlistat decreases endometrial cancer cell proliferation: implications for a novel treatment strategy

Abstract Objectives: Fatty acid synthase (FAS) is a key enzyme involved in fatty acid synthesis. It also plays an important role in tumorigenesis and is highly expressed in many cancers, including endometrial cancer. Orlistat is a weight loss medication that has been shown to be a potent inhibitor of FAS. The goal of this study was to evaluate the anti-tumorigenic potential of orlistat in endometrial cancer cell lines. Methods: The endometrial cancer cell lines ECC-1 and KLE were treated with varying doses of orlistat (0.01-500 μM). Cell proliferation was assessed using the MTT assay. Cell cycle progression was evaluated by Cellometer and apoptosis was assessed using the Annexin V assay. Reactive oxygen species (ROS) was measured using the DCFH-DA assay. Western immunoblotting was performed to determine changes in FAS, cellular stress, cell cycle progression and the AMPK/mTOR pathways. Results: Orlistat inhibited cell proliferation in a dose-dependent manner in both cell lines (IC50 = 81.7 μM in ECC-1; IC50 = 145.5 μM in KLE). Treatment with orlistat resulted in G1 arrest but did not affect apoptosis. Orlistat increased ROS at high doses (500 uM, p = 0.008 for ECC-1 cells and p = 0.012 for KLE cells) and induced expression of BIP and PERK. Western immunoblot analysis demonstrated that orlistat decreased expression of FAS, acetyl-CoA carboxylase (ACC), and carnitine palmitoyltransferase 1A (CPT1A), important proteins in fatty acid metabolism. Treatment with orlistat also increased expression of phosphorylated AMPK and decreased expression of the downstream S6 protein. Conclusions: Orlistat suppressed cell growth in endometrial cancer cell lines through inhibition of fatty acid metabolism, induction of cell cycle G1 arrest, and modulation of the AMPK/mTOR pathway. Given that patients with endometrial cancer have high rates of obesity, orlistat should be further investigated as a novel strategy for endometrial cancer treatment and prevention. Citation Format: Weiya Z. Wysham, Dario R. Roque, Jianjun Han, Hui Guo, Lu Zhang, Paola A. Gehrig, Chunxiao Zhou, Victoria L. Bae-Jump. Orlistat decreases endometrial cancer cell proliferation: implications for a novel treatment strategy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2994.

Effects of Fatty Acid Synthase Inhibition by Orlistat on Proliferation of Endometrial Cancer Cell Lines.

Fatty acid synthase (FAS) is a key lipogenic enzyme that is highly expressed in endometrial cancer. Orlistat is a weight loss medication that has been shown to be a potent inhibitor of FAS. The goal of this study was to evaluate the anti-tumorigenic potential of orlistat in endometrial cancer cell lines. The endometrial cancer cell lines ECC-1 and KLE were used. Cell proliferation was assessed by MTT assay after treatment with orlistat. Cell cycle progression was evaluated by Cellometer and apoptosis was assessed using the Annexin V assay. Reactive oxygen species (ROS) was measured using the DCFH-DA assay. Western immunoblotting was performed to determine changes in FAS, cellular stress, cell cycle progression, and the AMPK/mTOR pathways. Orlistat inhibited cell proliferation by 61% in ECC-1 cells and 57% in KLE cells at a dose of 500μM. Treatment with orlistat at this concentration resulted in G1 arrest (p < 0.05) but did not affect apoptosis. Orlistat increased ROS and induced the expression of BIP (1.28-fold in ECC-1 compared to control, p < 0.05; 1.92-fold in KLE, p < 0.05) and PERK (2.25-fold in ECC-1, 1.4-fold in KLE, p < 0.05). Western immunoblot analysis demonstrated that orlistat decreased expression of important proteins in fatty acid metabolism including FAS (67% in ECC-1, 15% in KLE), acetyl-CoA carboxylase (40% in ECC-1, 35% in KLE), and carnitine palmitoyltransferase 1A (CPT1A) (65% in ECC-1, 25% in KLE) in a dose-dependent manner. In addition, orlistat at a dose of 500μM increased expression of phosphorylated-AMPK (1.9-fold in ECC-1, p < 0.01; 1.5-fold in KLE, p < 0.05) and decreased expression of phosphorylated-Akt (25% in ECC-1, p < 0.05; 37% in KLE, p < 0.05) and phosphorylated-S6 (68% in ECC-1, 56% in KLE). Orlistat inhibits cell growth in endometrial cancer cell lines through inhibition of fatty acid metabolism, induction of cell cycle G1 arrest, activation of AMPK and inhibition of the mTOR pathway. Given that patients with endometrial cancer have high rates of obesity, orlistat should be further investigated as a novel strategy for endometrial cancer treatment.

Stem cell protein Piwil1 endowed endometrial cancer cells with stem-like properties via inducing epithelial-mesenchymal transition.

BackgroundStem cell protein Piwil1 functions as an oncogene in various tumor types. However, the exact function and mechanism of Piwil1 in endometrial cancer remains unclear.MethodsThe expression of Piwil1 and its relationships with clinicopathological factors were investigated using immunohistochemistry. Up- or down-regulation of Piwil1 were achieved by stable or transient transfection with plasmids or short hairpin RNA (shRNA). Effects of Piwil1 on cancer cells viability, invasion and migration were evaluated by MTT, plate colony formation, transwell assay and nude mouse tumor xenograft assay. The stem-like properties of endometrial cancer cells was detected by spheroid formation assay. Effects of Piwil1 on expression levels of target genes were detected by qRT-PCR, western blotting and Immunofluorescence.ResultsCompared with atypical hyperplasia and normal tissues, Piwil1 was much higher in endometrial carcinoma tissues. We found that Piwil1 expression was significantly correlated with FIGO stage, lymphovascular space involvement, lymph node metastasis and level of myometrial invasion. Overexpression of Piwil1 functioned to maintain stem-like characteristics, including enhancing tumor cell viability, migration, invasion and sphere-forming activity. Conversely, Piwil1 knockdown inhibited cell viability, migration, invasion, sphere-forming activity in vitro and tumor formation in xenograft model in vivo. Furthermore, study of the expression of epithelial and mesenchymal markers showed that Piwil1 was responsible for an EMT-like phenotype associated with an increase in mesenchymal markers and suppression of E-cadherin. Moreover, Piwil1 augmented expression levels of CD44 and ALDH1 expression, two known endometrial CSC markers, as well as other stemness-associated genes.ConclusionsOur results suggested that stem cell protein Piwil1 play important roles in regulating EMT and the acquisition of stem-like properties of endometrial cancer cells. Therefore, it indicated that Piwil1 may represent a promising target for developing a novel treatment strategy for endometrial cancer.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-015-1794-8) contains supplementary material, which is available to authorized users.

Simvastatin, an HMG-CoA reductase inhibitor, exhibits anti-metastatic and anti-tumorigenic effects in endometrial cancer

ObjectiveOur goal was to evaluate the effects of simvastatin on endometrial cancer cell lines and primary cultures of endometrial cancer cells. MethodsCell proliferation in the ECC-1 and Ishikawa endometrial cancer cell lines and primary cultures of endometrial cancer cells was assessed by MTT assay. Apoptosis and cell cycle were detected by Annexin V assay and propidium iodide staining, respectively. Reactive oxygen species and cell adhesion were assessed using ELISA assays. Invasion was analyzed using a transwell invasion assay. Mitochondrial DNA damage was confirmed using qPCR. The effects of simvastatin on the AKT/mTOR and MAPK pathways were determined by Western blotting. ResultsSimvastatin inhibited cell proliferation in a dose-dependent manner in both endometrial cancer cell lines and 5/8 primary cultures of endometrial cancer cells. Simvastatin treatment resulted in G1 cell cycle arrest, a reduction in the enzymatic activity of HMG-CoA, induction of apoptosis as well as DNA damage and cellular stress. Treatment with simvastatin resulted in inhibition of the MAPK pathway and exhibited differential effects on the AKT/mTOR pathway in the ECC-1 and Ishikawa cells. Minimal change in AKT phosphorylation was seen in both cell lines. An increase in phosphorylated S6 was seen in ECC-1 and a decrease was seen in Ishikawa. Treatment with simvastatin reduced cell adhesion and invasion (p<0.01) in both cell lines. ConclusionSimvastatin had significant anti-proliferative and anti-metastatic effects in endometrial cancer cells, possibly through modulation of the MAPK and AKT/mTOR pathways, suggesting that statins may be a promising treatment strategy for endometrial cancer.

Over-Expression of Platelet-Derived Growth Factor-D Promotes Tumor Growth and Invasion in Endometrial Cancer

The platelet-derived growth factor-D (PDGF-D) was demonstrated to be able to promote tumor growth and invasion in human malignancies. However, little is known about its roles in endometrial cancer. In the present study, we investigated the expression and functions of PDGF-D in human endometrial cancer. Alterations of PDGF-D mRNA and protein were determined by real time PCR, western blot and immunohistochemical staining. Up-regulation of PDGF-D was achieved by stably transfecting the pcDNA3-PDGF-D plasmids into ECC-1 cells; and knockdown of PDGF-D was achieved by transient transfection with siRNA-PDGF-D into Ishikawa cells. The MTT assay, colony formation assay and Transwell assay were used to detect the effects of PDGF-D on cellular proliferation and invasion. The xenograft assay was used to investigate the functions of PDGF-D in vivo. Compared to normal endometrium, more than 50% cancer samples showed over-expression of PDGF-D (p < 0.001), and high level of PDGF-D was correlated with late stage (p = 0.003), deep myometrium invasion (p < 0.001) and lympha vascular space invasion (p = 0.006). In vitro, over-expressing PDGF-D in ECC-1 cells significantly accelerated tumor growth and promoted cellular invasion by increasing the level of MMP2 and MMP9; while silencing PDGF-D in Ishikawa cells impaired cell proliferation and inhibited the invasion, through suppressing the expression of MMP2 and MMP9. Moreover, we also demonstrated that over-expressed PDGF-D could induce EMT and knockdown of PDGF-D blocked the EMT transition. Consistently, in xenografts assay, PDGF-D over-expression significantly promoted tumor growth and tumor weights. We demonstrated that PDGF-D was commonly over-expressed in endometrial cancer, which was associated with late stage deep myometrium invasion and lympha vascular space invasion. Both in vitro and in vivo experiments showed PDGF-D could promote tumor growth and invasion through up-regulating MMP2/9 and inducing EMT. Thus, we propose targeting PDGF-D to be a potent strategy for endometrial cancer treatment.

Recent advances in research on epigenetic alterations and clinical significance of para-aortic lymphadenectomy in endometrial cancer: an introduction

Endometrial cancer is the most frequent cancer of the female reproductive organs in industrialized countries. In 2012, the numbers of new cases and deaths from endometrial cancer in the US were estimated to be 47,130 and 8,010, respectively [1]. The incidence of endometrial cancer is also increasing steadily in Japan, where the estimated number of new cases in 2007 was 9,104 [2] and the number of deaths in 2011 was 2,034 [3]. Endometrial cancer is a surgically staged disease and post-operative therapy is offered to patients with a high risk of recurrence according to the extent and aggressiveness of the tumor. Current topics in endometrial cancer include: the therapeutic significance of lymphadenectomy, the role of epigenetic alterations, and revision of the International Federation of Gynecology and Obstetrics staging criteria (FIGO 2008) for this disease. There was a paradigm shift in the treatment strategy for endometrial cancer after the introduction of a surgical staging system (FIGO 1988) that replaced the older clinical staging system. The newer paradigms of extended-surgical staging containing lymphadenectomy with more restricted use of adjuvant therapy and the older paradigm of simple hysterectomy bilateral salpingo-oophorectomy with more frequent use of adjuvant radiotherapy need to be compared prospectively in terms of survival benefits, quality of life, and cost of treatment [4]. Several issues regarding surgical staging need to be clarified. They include: how should suitable patients for complete lymphadenectomy be selected and what is the optimal extent of lymphadenenctomy? The therapeutic significance of lymphadenectomy has long been a matter of great debate. In 1964, Lewis suggested a therapeutic effect of pelvic lymphadenectomy in nodepositive patients [5]. He employed pelvic lymphadenectomy because endometrial cancer often recurred at the pelvic side wall after conventional hysterectomy and bilateral salpingo-oophorectomy, which suggested inadequate primary surgery. Retrospective studies suggest a therapeutic significance for lymphadenectomy, which is a function of removed lymph node count (thoroughness) and area of dissection (pelvic only versus pelvic and para-aortic lymphadenectomy) [6–8]. However, two prospective randomized controlled trials (RCTs) that intended to prove the therapeutic role of pelvic lymphadenectomy failed to show any survival advantage of pelvic lymphadenectomy versus no lymphadenectomy [9, 10]. However, there has been some criticism about the design of these trials because para-aortic lymphadenectomy was not included in the study arm. A retrospective cohort study which compared pelvic lymphadenectomy with combined pelvic and paraaortic lymphadenectomy revealed survival improvement in the pelvic and para-aortic lymphadenectomy group if this treatment was offered to intermediate-/high-risk endometrial cancer patients [11]. Based on these findings, discussions have begun about the design of future clinical trials to validate the therapeutic significance of lymphadenectomy. Topics for discussion include the eligibility of patients (all patients or selected patients at some risk of nodal metastasis), extent of lymphadenectomy (area: pelvic alone versus pelvic plus para-aortic, thoroughness: number of nodes removed), and type of experimental design (RCT versus cohort study). The difficulties and pitfalls of RCTs for validating surgical procedures have often been addressed [12–16]. These include the participating surgeons’ expertise in experimental N. Sakuragi (&) Department of Obstetrics and Gynecology, Hokkaido University School of Medicine, North 15, West 7, Kitaku, Sapporo 060-8638, Japan e-mail: sakuragi@med.hokudai.ac.jp

Current treatment strategies for endometrial cancer

Treatment paradigms for endometrial cancer are rapidly evolving. New minimally invasive surgical techniques have helped to reduce the morbidity of women with early-stage tumors undergoing hysterectomy. A growing body of evidence has helped to stratify women with intermediate-risk endometrial cancer to optimize adjuvant therapy in this subset of patients. For women with high-risk and metastatic endometrial cancer there has been a renewed interest in incorporating chemotherapy into treatment paradigms. This review summarizes the latest advances in the treatment of endometrial cancer with a special focus on adjuvant therapy.

Regional differences in endometrial cancer treatment and outcomes: A provincial study

5095 Background: The optimal treatment strategy for endometrial cancer is controversial, and there is lack of treatment standardization across the province of Ontario. The objective of this study is to compare endometrial cancer treatment and outcomes among five centres in Ontario, Canada. Methods: This was a retrospective cohort study of women diagnosed with endometrial cancer in Ontario between 1996 and 1998. Approximately 30–50 patient charts were randomly selected by medical records at each of five health care centres in Ontario. Study outcomes included overall and disease-free survival, rates of adjuvant radiotherapy, complications of treatment, and predictive factors for survival. Results: There were 195 women in this cohort. The two different treatment strategies were total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH BSO) and surgical staging (TAH BSO and pelvic lymph node dissection). There was a wide range of staging practices across the province; none of the patients in Kings...