Abstract
BackgroundTo date, tools to study metastasis in endometrial cancers are insufficiently developed. The aim of this study was to characterize the cell line EN-1078D, a new endometrial carcinoma cell line derived from a metastasis to the ovary.Methods and ResultsCells were characterized using cytology, transmission electron microscopy, karyotyping and morphological appearance in culture. Molecular features were determined by RT-PCR, Western Blot, FISH and sequencing. MTT proliferation assays were performed to investigate the sensitivity of EN-1078D to anticancer agents such as cisplatin and doxorubicin. Also, subcutaneous and intravenous injections in nude mice were done to test the tumorigenic and metastatic properties of EN-1078D cells. Our results indicate that EN-1078D cells express both oestrogen receptors isoforms (ER alpha and ER beta) and also low levels of progesterone receptor B (PR-B). In addition, this cell line expresses high levels of MMP-2 and MMP-14 mRNA, low levels of TIMP-1 and TIMP-2 transcripts and no detectable levels of MMP-9 mRNA. Moreover, all nude mice developed tumors by subcutaneous injections and cell invasion was observed in vitro in response to TGF-beta 3. Her-2/neu was not overamplified but mutations in the C-2 domain of PTEN gene as well as codon 12 of the K-Ras gene were found. Finally, EN-1078D shows sensitivity to drugs commonly used in chemotherapy such as cisplatin and doxorubicin: IC50 of 2.8 μM of cisplatin after 72 hours of exposure and 0.54 μM of doxorubicin after 48 hours.ConclusionTaken together, these results suggest that EN-1078D will be an excellent tool to study the properties of metastatic endometrial cancer cells in vitro and their regulation by sex steroids.
Highlights
To date, tools to study metastasis in endometrial cancers are insufficiently developed
HeLa, a cervical cancer cell line, which we have previously found to be sensitive to these drugs was used as positive control, while KLE, an endometrial adenocarcinoma cell line poorly inhibited by the drug, was used as negative control
Two other cell lines were used as a reference: HeLa, a cervical cancer cell line that we showed to be sensitive to these anticancer agents, and KLE, an endometrial cancer cell line known to be chemoresistant [28]
Summary
Tools to study metastasis in endometrial cancers are insufficiently developed. The aim of this study was to characterize the cell line EN-1078D, a new endometrial carcinoma cell line derived from a metastasis to the ovary. Ninety-seven percent of all cancers of the uterus arise from the glands of the endometrium and are known as endometrial carcinomas [1]. When diagnosed at early stages of the disease, this type of cancer is a highly curable malignancy with a 5-year relative survival rate of more than 80% [2]. Patients presenting metastases have a 5-year survival rate of less than 20% [2]. Metastasis represents the main cause of death for patients with endometrial carcinoma. Very few models are available, to date, for the experimental characterization of factors involved in the metastatic phenotype in endometrial carcinoma cells
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