Tumor Progression In Endometrial Cancer Research Articles

T-Box Transcription Factor 2 Mediates Chemoresistance of Endometrial Cancer via Regulating FSP1-involved Ferroptosis.

Chemotherapy is increasingly being used in the first-line treatment of endometrial cancer (EC) patients. However, chemoresistance seriously affects its efficacy. Understanding the underlying molecular mechanisms is critical for EC treatment. We explored the regulatory role of T-Box transcription factor 2 (TBX2)-ferroptosis suppressor protein 1 (FSP1) axis in ferroptosis and chemoresistance of EC. Cisplatin-resistant cell line Ishikawa/DDP cells were utilized to generate TBX2 and FSP1 overexpression and knockdown stable cell lines by using lentivirus infection and puromycin selection. Cell viability and ferroptosis status were evaluated in EC cells with or without Cisplatin and/or FSP1 inhibitor (iFSP1) using CKK-8, lipid peroxidation, malondialdehyde, and lactate dehydrogenase release assays. Endometrial carcinoma xenograft mouse model was established to further explore the function of TBX2-FSP1 axis on ferroptosis and tumor progression in EC. TBX2 suppressed Cisplatin-induced ferroptosis through up-regulating FSP1 expression level in EC cells. On the contrary, knockdown of TBX2 reduced FSP1 expression and significantly promoted Cisplatin-induced ferroptosis. TBX2 or FSP1 overexpression and knockdown promote and inhibit EC tumor growth under Cisplatin treatment, respectively. Interestingly, silence FSP1 could reverse TBX2-mediated ferroptosis inhibition and tumor-promoting effect. TBX2-FSP1 axis inhibits ferroptosis and enhances the Cisplatin resistance, which will provide an important theoretical basis and potential solution for the clinical treatment of EC.

The prognostic role of mitosis index, stage and grade of endometrial cancer in Dr. Soetomo General Academic Hospital Surabaya, Indonesia, in 2018-2020

HIGHLIGHTS 1. Mitotic index and grade are prognostic factors for endometrial cancer, but both are independent.2. Stage and mitotic index associated with cell proliferation affect the prognosis of endometrial cancer. ABSTRACT Objective: This study aimed to analyze the correlation between the mitotic index and the stage and grade of endometrial cancer. Materials and Methods: We collected pathology reports of endometrial cancer from the Pathology Laboratory at Dr. Soetomo General Hospital in Surabaya, Indonesia, covering cases diagnosed between 2018 and 2020. A total of 106 cases of endometrial cancer were included in this study. For each case, detailed records of the cancer stage, grade, and mitotic index were recorded. The mitotic index, an indicator of cell proliferation, was quantified, and its correlation with cancer stage and grade was assessed. To determine the strength and direction of these relationships, we performed a Spearman rank correlation statistical analysis for non-parametric data. Results: Our findings indicated a significant positive correlation between the mitotic index and the stage of endometrial cancer. An increase in the mitotic index, reflecting a higher proliferation rate of cancer cells, was associated with a more advanced cancer stage, suggesting that the mitotic index could potentially serve as a prognostic marker for assessing tumor progression in endometrial cancer. However, our analysis revealed no significant correlation between the mitotic index and the histological grade of endometrial cancer, implying that the grade, which typically reflects the differentiation status and morphological characteristics of the tumor cells, is independent of the proliferation rate as measured by the mitotic index. Conclusion: The mitotic index is positively correlated with the stage of endometrial cancer but does not show a correlation with the histological grade. These findings highlight the potential use of the mitotic index in staging endometrial cancer.

CENPA promotes glutamine metabolism and tumor progression by up-regulating SLC38A1 in endometrial cancer

Glutamine addiction is a significant hallmark of metabolic reprogramming in tumors and is crucial to the progression of cancer. Nevertheless, the regulatory mechanisms of glutamine metabolism in endometrial cancer (EC) remains elusive. In this research, we found that elevated expression of CENPA and solute carrier family 38 member 1 (SLC38A1) were firmly associated with worse clinical stage and unfavorable outcomes in EC patients. In addition, ectopic overexpression or silencing of CENPA could either enhance or diminish glutamine metabolism and tumor progression in EC. Mechanistically, CENPA directly regulated the transcriptional activity of the target gene, SLC38A1, leading to enhanced glutamine uptake and metabolism, thereby promoting EC progression. Notably, a prognostic model utilizing the expression levels of CENPA and SLC38A1 genes independently emerged as a prognostic factor for EC. More importantly, CENPA and SLC38A1 were significantly elevated and positively correlated, as well as indicative of poor prognosis in multiple cancers. In brief, our study confirmed that CENPA is a critical transcription factor involved in glutamine metabolism and tumor progression through modulating SLC38A1. This revelation suggests that targeting CENPA could be an appealing therapeutic approach to address pan-cancer glutamine addiction.

Evaluation of serum levels of soluble (s)L- and (s)P-selectins in endometrial cancer.

A number of reports on the role of selectin in the process of carcinogenesis, at the stage of proliferation and metastasis, have been available. The aim of the study was to analyze (s)P- and (s)L-selectin serum concentrations in women with EC and to compare these concentrations to clinical/pathological parameters and disease progression using surgical-pathological staging data. A total of 46 patients with EC and 50 healthy controls were included in the study. Serum concentrations of sL- and sP-selectins were measured in all participants. The oncologic protocol was implemented in all women from the study group. Significantly higher serum concentrations were found in EC women as compared to controls. No statistically significant differences were found between the concentrations of the soluble forms of selectins and the following parameters: histologic type of EC, histologic tumor differentiation, depth of myometrial infiltration, cervical involvement, distant metastases, vascular space invasion, and disease advancement. Slightly higher (s)P-selectin concentrations were observed in serous carcinoma, in women with cervical involvement, in the sera of women with vascular space invasion and with advanced stages of the disease. Slightly higher mean (s)P-selectin concentrations correlated with lower differentiation of the tumor. Slightly higher mean (s)P-selectin concentration was detected in the sera of women with lymph node metastases and with the serosal and/or adnexal involvement. The results were statistically insignificant, but they almost reached statistical significance. L- and P-selectins play a role in the biology of EC. The absence of an unambiguous relationship between differences in (s)L- and (s)P-selectin levels and disease advancement suggests that they do not play a vital role in tumor progression in endometrial cancer.

Estrogen-related receptor alpha induces epithelial-mesenchymal transition through cancer-stromal interactions in endometrial cancer

Estrogen-related receptor alpha (ERRα), which shares structural similarities with estrogen receptors, is associated with tumor progression in endometrial cancer, but little is known about the detailed underlying mechanism. We investigated whether ERRα, in cooperation with peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), could participate in epithelial-mesenchymal transition (EMT) in endometrial cancer through cancer-stromal interactions. Two endometrial cancer cell lines, Ishikawa and HEC-1A, transfected with ERRα/PGC-1α expression plasmids or silenced for ERRα expression, were co-cultured with telomerase-transformed human endometrial stromal cells (T-HESCs). We found that EMT-associated factors including vimentin, Snail, and zinc finger E-box binding homeobox 1 were upregulated in cancer cells overexpressing ERRα/PGC-1α and that transforming growth factor-beta (TGF-β) was induced in T-HESCs in the same conditions. In contrast, ERRα knockdown suppressed EMT-associated factors in cancer cells and TGF-β in T-HESCs. ERRα/PGC-1α overexpression increased the expression of EMT-associated factors after TGF-β exposure; however, it decreased E-cadherin at protein level. ERRα knockdown suppressed EMT-associated factors in the presence of TGF-β, whereas E-cadherin remained unchanged. Matrigel invasion assays revealed that ERRα knockdown attenuated the stimulation of migration and invasion by TGF-β. These findings suggest that ERRα is a potential target for inhibiting TGF-β-induced EMT through cancer-stromal interactions in endometrial cancer.

RIZ1 is regulated by estrogen and suppresses tumor progression in endometrial cancer

Endometrial cancer (EC) is the estrogen-dependent gynecologic malignancy, however the molecular mechanism involved in the development and progression of EC remain unclear. The aim of this study was to investigate the role of RIZ1 in EC. Immunohistochemical analysis revealed that RIZ1was decreased in EC than in normal endometrium. Lower RIZ1 level was correlated with high-grade carcinoma (p = 0.048) and positive expression of ERα (p = 0.004). In EC cells, estrogen could down regulated the expression of RIZ1, however, ICI182,780 could up regulated the expression of RIZ1. Besides, in vitro and in vivo, RIZ1 could remarkably suppress tumor proliferation, metastasis and invasion. Our data support that RIZ1 was a novel tumor suppressor and could provide a potential therapeutic target in human EC.

Rap1GAP inhibits tumor progression in endometrial cancer

ObjectiveEndometrioid adenocarcinoma (EAC) is a common endometrial cancer with recent dramatic increases in incidence. Previous findings indicate that Rap1GAP acts as a tumor suppressor inhibiting Ras superfamily protein Rap1 in multiple aggressive carcinomas; however, Rap1GAP expression in EAC has not been investigated. In this study, the tumor suppressing activity of Rap1GAP in EAC was explored. MethodsEAC cell lines were used to examine Rap1GAP levels by real-time RT-PCR and western blotting and the effects of Rap1GAP on cancer cell invasion and migration. Rap1GAP expression was analyzed by immunohistochemical staining for Rap1GAP, E-cadherin in surgically resected tumors of 114 EAC patients scored according to EAC differentiation grade. Prognostic variables such as age, stage, grade, tumor size, and immunostaining for Rap1GAP, E-cadherin were evaluated using Cox regression multivariate analysis. ResultsLow Rap1GAP expression was detected in poorly differentiated EAC cells. Rap1GAP deficiency significantly accelerated while Rap1 deficiency decreased cancer cell migration and invasion. Patients with higher Rap1GAP, E-cadherin, and especially combined Rap1GAP/E-cadherin levels had better overall survival than EAC patients with no or weak expression. In addition, Rap1GAP expression was an independent prognostic factor in EAC. ConclusionsInhibition of Rap1GAP expression increases EAC cell migration and invasion through upregulation of Rap1. Low expression of Rap1GAP correlates with poor EAC differentiation. Our findings suggest that Rap1GAP is an important tumor suppressor with high prognostic value in EAC.

MiR-944 acts as a prognostic marker and promotes the tumor progression in endometrial cancer.

microRNA-944 (miR-944) has been reported to be deregulation and play either tumor suppressive or oncogenic function in human malignancies. Previous studies have reported that miR-944 is overexpressed in gynecological malignancies including cervical cancer and breast cancer. While, the clinical significance of miR-944 and its function in human endometrial carcinoma (EC) keep unknown. The levels of miR-944 were analyzed in 68 EC tissues and 20 normal endometrial tissues. Results showed that miR-944 was significantly overexpressed in EC tissues compared to normal endometrial tissues. In addition, increased levels of miR-944 also observed in EC cell lines. Clinicopathological analysis verified that miR-944 expression was associated with international federation of gynecology and obstetrics (FIGO) stages and pathology classification of EC. Moreover, Kaplan-Meier analysis found that miR-944 highly expressing EC patients showed a notable shorter survival. Further experiments revealed that miR-944 knockdown significantly inhibited growth and cell cycle progression while facilitated apoptosis in Ishikawa cells. In turn, miR-944 overexpression prominently promoted proliferation and cell cycle progression, and suppressed apoptosis in KLE cells. In vivo experiments further confirmed that miR-944 silencing notably restrained the tumor growth of EC in nude mice. Mechanically, cell adhesion molecule 2 (CADM2) was recognized as a direct downstream target of miR-944 in EC cells. An inverse correlation between miR-944 and CADM2 expression was observed in EC tissues. Interestingly, CADM2 overexpression showed similar effects to miR-944 knockdown in Ishikawa cells with decreased growth, cell cycle arrest at G1 phase and increased apoptosis. Taken together, this work support the first evidence that miR-944 can be potentially used as a promising biomarker and novel therapeutic target for human EC.

Abstract 60: Mig-6 suppresses development and progression of endometrial cancer by inhibiting ERK2 phosphorylation

Abstract Endometrial cancer is the most common cancer of the female reproductive system. PTEN is mutated or absent in more than half of human endometrial cancers. The major pathologic phenomenon of endometrial cancer is the loss of ovarian steroid hormone control over uterine epithelial cell proliferation and apoptosis. Mig-6 suppresses estrogen signaling. Here, we show the significance of MIG-6 in human endometrial cancer through sample analysis, where MIG-6 expression is inversely associated with ERK phosphorylation; this relationship is tightly correlated with endometrial cancer progression. To determine the tumor suppressor function of Mig-6 in the development of endometrial cancer, we generated Mig-6 conditional overexpression mice (R26Mig-6LSL). To assess the effects of Mig-6 on the PTEN/PI3K/AKT signaling pathway in uterine tumorigenesis, mice with Pten floxed (Ptenf/f) and R26Mig-6LSL were bred to the PRCre mouse model to generate overexpression of Mig-6 and ablation of Pten in the uterus (PRcre/+ R26Mig-6LSL Ptenf/f). PRcre/+ R26Mig-6LSL Ptenf/f showed significantly increased survival time and uterine weight compared to PRcre/+ Ptenf/f mice. Gross morphology and histological analysis displayed dramatically suppressed development of endometrial cancer in double mutant mice compared to ablation of Pten alone. Immunohistochemical analysis showed significantly increased apoptosis and decreased proliferation in epithelial cells of PRcre/+ R26Mig-6LSL Ptenf/f mice compared to PRcre/+ Ptenf/f mice. Interestingly, the expression of pERK1/2 was significantly decreased in PRcre/+ R26Mig-6LSL Ptenf/f mice compared to PRcre/+ Ptenf/f mice. To examine whether inhibition of ERK phosphorylation suppresses tumor progression in endometrial cancer, PRcre/+Ptenf/f mice were treated with U0126, an effective inhibitor of MAPK/ERK kinase. PRcre/+Ptenf/f mice treated with U0126 exhibited a significant reduction in uterine weight. Histopathological analysis of the entire animal cohort showed that inhibition of ERK phosphorylation suppressed endometrial cancer progression in PRcre/+ Ptenf/f mice, as reflected by the arrest of tumors at the hyperplastic or normal stage, whereas tumors from PRcre/+Ptenf/f mice treated with vehicle advanced to endometrial cancer. These results demonstrate that activation of ERK signaling is critical for endometrial cancer development and progression in Pten mutation. Our findings highlight a crucial tumor suppressor role for MIG-6 in progression of PTEN-null endometrial cancer by inhibiting ERK phosphorylation. As MIG-6 is a mediator of progesterone signaling, the activity of which can suppress unopposed-estrogen signaling, our studies provide a potential new drug target for the intervention of metastatic human endometrial cancer. (This work was supported by NIH, U54 HD007495 to J.P.L, NIH, R01 HD057873 and American Cancer Society Research Grant, RSG-12-084-01-TBG to J.W.J.) Citation Format: Jung-Yoon Yoo, Tae Hoon Kim, Hong Im Kim, Jane Li, Gordon B. Mills, Russell R. Broaddus, John P. Lydon, Ho-Geun Yoon, Jae-Wook Jeong. Mig-6 suppresses development and progression of endometrial cancer by inhibiting ERK2 phosphorylation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 60. doi:10.1158/1538-7445.AM2014-60

HE4 (WFDC2) Promotes Tumor Growth in Endometrial Cancer Cell Lines

HE4, also known as WFDC2, is a useful biomarker for ovarian cancer when either used alone or in combination with CA125. HE4 is also overexpressed in endometrial cancer (EC), but its function in cancer cells is not clear. In this study, we investigate the role of HE4 in EC progression. An HE4-overexpression system was established by cloning the HE4 prototypic mRNA variant (HE4-V0) into a eukaryotic expression vector. Following transfection, stable clones in two EC cell lines were selected. The effects of HE4 overexpression on cell growth and function were measured with the use of cell proliferation assay, matrigel invasion, and soft agar gel colony formation assays. HE4-induced cancer cell proliferation in vivo was examined in a mouse xenograft model. HE4 overexpression significantly enhanced EC cell proliferation, matrigel invasion, and colony formation in soft agar. Moreover, HE4 overexpression promoted tumor growth in the mouse xenograft model. HE4 overexpression enhanced several malignant phenotypes in cell culture and in a mouse model. These results are consistent with our previous observation that high levels of serum HE4 closely correlate with the stage, myometrial invasion and tumor size in patients with EC. This study provides evidence that HE4 overexpression directly impacts tumor progression in endometrial cancer.

Expression of Astrocyte Elevated Gene-1

To assess the expression of astrocyte elevated gene-1 (AEG-1) in different endometrial specimens and to investigate its relationship with clinicopathological features and its impact on patient outcome. By Western blot analysis, we investigated AEG-1 expression in paired endometrial tissues and adjacent nontumor tissues of the same patients (n = 4). Immunohistochemistry analysis was used to determine the expression levels of AEG-1 protein in 35 normal endometrium, 40 atypical endometrial hyperplasia, and 174 endometrial cancers. The correlation between AEG-1 expression and various clinicopathological characteristics of endometrial cancer patients was analyzed. Western blot analysis showed that AEG-1 expression levels were up-regulated in all 4 human primary endometrial cancer tissues compared with their matched adjacent noncancerous tissues. The frequent and strong expression of AEG-1 was gradually elevated in normal endometrial tissue, atypical hyperplasia, and endometrial cancers (P < 0.001). Although AEG-1 staining mainly emerged in the cytoplasm, a nuclear distribution was observed in both invasive and advanced endometrial cancer cells. The Ki67 (a proliferation marker) was frequently expressed in the high AEG-1-expressed area, whereas areas with low AEG-1 levels showed weak Ki67 expression. Astrocyte elevated gene-1 overexpression was positively correlated with the International Federation of Gynecology and Obstetrics stage, depth of myometrial invasion, lymph node metastasis, lymph vascular space invasion, and recurrence but not with age or histological type. Patients with high AEG-1 expression had significantly poor overall survival and disease-free survival compared with patients with AEG-1 low expression (both P < 0.001). Multivariate Cox proportional hazards regression demonstrated that high AEG-1 expression was an independent prognostic factor for both the overall survival and disease-free survival of patients with endometrial cancer (P = 0.002 and P = 0.004, respectively). Astrocyte elevated gene-1 overexpression may be associated with carcinogenesis and tumor progression in endometrial cancer. Moreover, it may be a new prognostic marker or a target for improving the treatment efficiency of patients with endometrial cancer.

Prognostic significance of stromal versican expression in human endometrial cancer

Versican expression may enhance tumor invasion and metastasis. However, the expression of versican in human endometrial cancer has seldom been characterized. The aim of this study was to investigate versican expression in endometrial cancers. We immunohistochemically investigated the expression of versican protein in 167 endometrial cancers and analyzed the correlation with various observed clinicopathological features, including patient outcome. Stromal versican expression was significantly higher in the advanced-stage (P = 0.010) and high-grade (P = 0.049) cancers, lymph node metastasis (P = 0.012), and ovarian metastasis (P = 0.024). Epithelial versican expression was significantly higher in patients with lymph node metastasis (P = 0.014) and lymph-vascular space involvement (P = 0.014). The disease-free survival (DFS) and overall survival (OS) rates of patients exhibiting high stromal versican expression were significantly lower than those of patients exhibiting low stromal versican expression (P < 0.0001). Multivariate analysis showed that high stromal versican expression was an independent prognostic factor for DFS and OS. Versican enrichment of the stroma may be associated with tumor progression in endometrial cancer. Stromal versican expression can serve as an indicator of poor prognosis for patients with endometrial cancer.

Expression pattern of the AP-1 family in endometrial cancer: correlations with cell cycle regulators.

To study the expression pattern and the role of the AP-1 (activating protein-1) family of transcription factors in endometrial carcinogenesis. We performed Western blot experiments with specific antibodies for each of the AP-1 proteins (c-jun, junB, junD, c-fos, fosB, fra-1, fra-2) with 41 endometrial carcinomas. Expression levels of the AP-1 factors were correlated with clinico-pathologic tumor parameters, steroid receptor status, Ki-67 expression and the expression levels of eight cell cycle regulatory proteins (cyclin D1, cyclin E, cyclin B1, cdk2, cdk4, p16, p21, and Rb). Of the seven AP-1 factors, three (c-fos, fra2, and junB) clearly showed higher expression levels in tumors when compared to matched normal endometrial samples. These factors also correlated significantly with cell cycle promoters (c-fos with cyclin E, cyclin B1, cdk2, and cdk4; fra-2 with cyclin B1; and junB with cyclin D1). Furthermore, high c-fos expression correlated with low ER and PR immunoreactivity and high grading (G3). On the other hand, correlations with classic cell cycle inhibitors (Rb, p16, p21) have also been observed for all AP-1 factors except c-jun and junD. Our results indicate that the AP-1 family of transcription factors is probably implicated in the regulation of cell cycle progression and control in endometrial carcinomas. In particular, c-fos might be an additional negative prognostic factor and/or implicated in tumor progression in endometrial cancer.