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Reproductive Biology and Endocrinology
Published in: Reproductive Biology and Endocrinology 1/2007

Open Access 01-12-2007 | Research

Characterization of EN-1078D, a poorly differentiated human endometrial carcinoma cell line: a novel tool to study endometrial invasion in vitro

Authors: Marie-Claude Dery, Celine Van Themsche, Diane Provencher, Anne-Marie Mes-Masson, Eric Asselin

Published in: Reproductive Biology and Endocrinology | Issue 1/2007

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Abstract

Background

To 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 Results

Cells 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.

Conclusion

Taken 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.
Appendix
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Literature
1.
2.
Jemal A, Tiwari RC, Murray T, Ghafoor A, Samuels A, Ward E, Feuer EJ, Thun MJ: Cancer statistics, 2004. CA Cancer J Clin. 2004, 54: 8-29.CrossRefPubMed
3.
Cooper D, Schermer A, Sun TT: Classification of human epithelia and their neoplasms using monoclonal antibodies to keratins: strategies, applications, and limitations. Lab Invest. 1985, 52: 243-256.PubMed
4.
Omary MB, Ku NO, Liao J, Price D: Keratin modifications and solubility properties in epithelial cells and in vitro. Subcell Biochem. 1998, 31:105-40.: 105-140.
5.
Markman M: Hormonal therapy of endometrial cancer. Eur J Cancer. 2005, 41: 673-675. 10.1016/j.ejca.2004.12.008.CrossRefPubMed
6.
Kato H, Kato S, Kumabe T, Sonoda Y, Yoshimoto T, Kato S, Han SY, Suzuki T, Shibata H, Kanamaru R, Ishioka C: Functional evaluation of p53 and PTEN gene mutations in gliomas. Clin Cancer Res. 2000, 6: 3937-3943.PubMed
7.
Fujimoto J, Sakaguchi H, Aoki I, Khatun S, Toyoki H, Tamaya T: Steroid receptors and metastatic potential in endometrial cancers. J Steroid Biochem Mol Biol. 2000, 75: 209-212. 10.1016/S0960-0760(00)00176-X.CrossRefPubMed
8.
Vegeto E, Shahbaz MM, Wen DX, Goldman ME, O'Malley BW, McDonnell DP: Human progesterone receptor A form is a cell- and promoter-specific repressor of human progesterone receptor B function. Mol Endocrinol. 1993, 7: 1244-1255. 10.1210/me.7.10.1244.PubMed
9.
Curry TE, Osteen KG: The matrix metalloproteinase system: changes, regulation, and impact throughout the ovarian and uterine reproductive cycle. Endocr Rev. 2003, 24: 428-465. 10.1210/er.2002-0005.CrossRefPubMed
10.
Goldman S, Shalev E: The role of the matrix metalloproteinases in human endometrial and ovarian cycles. Eur J Obstet Gynecol Reprod Biol. 2003, 111: 109-121. 10.1016/S0301-2115(03)00341-5.CrossRefPubMed
11.
Nguyen M, Arkell J, Jackson CJ: Human endothelial gelatinases and angiogenesis. Int J Biochem Cell Biol. 2001, 33: 960-970. 10.1016/S1357-2725(01)00007-3.CrossRefPubMed
12.
Aglund K, Rauvala M, Puistola U, Angstrom T, Turpeenniemi-Hujanen T, Zackrisson B, Stendahl U: Gelatinases A and B (MMP-2 and MMP-9) in endometrial cancer-MMP-9 correlates to the grade and the stage. Gynecol Oncol. 2004, 94: 699-704. 10.1016/j.ygyno.2004.06.028.CrossRefPubMed
13.
Risinger JI, Hayes AK, Berchuck A, Barrett JC: PTEN/MMAC1 mutations in endometrial cancers. Cancer Res. 1997, 57: 4736-4738.PubMed
14.
Risinger JIFAU, Maxwell GLFAU, Chandramouli GVFAU, Jazaeri AFAU, Aprelikova OFAU, Patterson TFAU, Berchuck AFAU, Barrett JC: Microarray analysis reveals distinct gene expression profiles among different histologic types of endometrial cancer.
15.
Tashiro H, Blazes MS, Wu R, Cho KR, Bose S, Wang SI, Li J, Parsons R, Ellenson LH: Mutations in PTEN are frequent in endometrial carcinoma but rare in other common gynecological malignancies. Cancer Res. 1997, 57: 3935-3940.PubMed
16.
Levine RL, Cargile CB, Blazes MS, van Rees B, Kurman RJ, Ellenson LH: PTEN mutations and microsatellite instability in complex atypical hyperplasia, a precursor lesion to uterine endometrioid carcinoma. Cancer Res. 1998, 58: 3254-3258.PubMed
17.
Sherman ME: Theories of endometrial carcinogenesis: a multidisciplinary approach. Mod Pathol. 2000, 13: 295-308. 10.1038/modpathol.3880051.CrossRefPubMed
18.
Graziano SL, Gamble GP, Newman NB, Abbott LZ, Rooney M, Mookherjee S, Lamb ML, Kohman LJ, Poiesz BJ: Prognostic significance of K-ras codon 12 mutations in patients with resected stage I and II non-small-cell lung cancer. J Clin Oncol. 1999, 17: 668-675.PubMed
19.
Choudhury A, Kiessling R: Her-2/neu as a paradigm of a tumor-specific target for therapy. Breast Dis. 2004, 20:25-31.: 25-31.
20.
Hetzel DJ, Wilson TO, Keeney GL, Roche PC, Cha SS, Podratz KC: HER-2/neu expression: a major prognostic factor in endometrial cancer. Gynecol Oncol. 1992, 47: 179-185. 10.1016/0090-8258(92)90103-P.CrossRefPubMed
21.
Lazar E, Tudose N, Lazar DC: Prognostic significance of c-erbB2 protein in endometrial cancer. Rom J Morphol Embryol. 1998, 44: 101-107.PubMed
22.
Rolitsky CD, Theil KS, McGaughy VR, Copeland LJ, Niemann TH: HER-2/neu amplification and overexpression in endometrial carcinoma. Int J Gynecol Pathol. 1999, 18: 138-143.CrossRefPubMed
23.
Lounis H, Provencher D, Godbout C, Fink D, Milot MJ, Mes-Masson AM: Primary cultures of normal and tumoral human ovarian epithelium: a powerful tool for basic molecular studies. Exp Cell Res. 1994, 215: 303-309. 10.1006/excr.1994.1346.CrossRefPubMed
24.
Mobus VJ, Gerharz CD, Weikel W, Merk O, Dreher L, Kreienberg R, Moll R: Characterization of a human carcinosarcoma cell line of the ovary established after in vivo change of histologic differentiation. Gynecol Oncol. 2001, 83: 523-532. 10.1006/gyno.2001.6425.CrossRefPubMed
25.
Yamada T, Ueda M, Maeda T, Okamoto Y, Miyawaki Y, Otsuki Y, Ueki M, Sugimoto O: Establishment and characterization of CA 125 producing cell line (OMC-2) originating from a human endometrial adenocarcinoma. Asia Oceania J Obstet Gynaecol. 1989, 15: 403-416.CrossRefPubMed
26.
Geran RI GNH: Protocols for screening chemical agents and natural products against animal tumors and other biological systems. Third Edition. 1972, National Cancer Institute, 1-87.
27.
Cianciulli AM, Guadagni F, Marzano R, Benevolo M, Merola R, Giannarelli D, Marandino F, Vocaturo G, Mariani L, Mottolese M: HER-2/neu oncogene amplification and chromosome 17 aneusomy in endometrial carcinoma: correlation with oncoprotein expression and conventional pathological parameters. J Exp Clin Cancer Res. 2003, 22: 265-271.PubMed
28.
Gagnon V, Mathieu I, Sexton E, Leblanc K, Asselin E: AKT involvement in cisplatin chemoresistance of human uterine cancer cells. Gynecol Oncol. 2004, 94: 785-795. 10.1016/j.ygyno.2004.06.023.CrossRefPubMed
29.
Van Themsche C, Mathieu I, Parent S, Asselin E: Transforming growth factor-beta3 increases the invasiveness of endometrial carcinoma cells through phosphatidylinositol 3-kinase-dependent up-regulation of X-linked inhibitor of apoptosis and protein kinase c-dependent induction of matrix metalloproteinase-9. J Biol Chem. 2007, 282: 4794-4802. 10.1074/jbc.M608497200.CrossRefPubMed
30.
Kuramoto H, Tamura S, Notake Y: Establishment of a cell line of human endometrial adenocarcinoma in vitro. Am J Obstet Gynecol. 1972, 114: 1012-1019.PubMed
31.
32.
Way DL, Grosso DS, Davis JR, Surwit EA, Christian CD: Characterization of a new human endometrial carcinoma (RL95-2) established in tissue culture. In Vitro. 1983, 19: 147-158. 10.1007/BF02618053.CrossRefPubMed
33.
DAWE CJ, BANFIELD WG, MORGAN WD, SLATICK MS, CURTH HO: GROWTH IN CONTINUOUS CULTURE, AND IN HAMSTERS, OF CELLS FROM A NEOPLASM ASSOCIATED WITH ACANTHOSIS NIGRICANS. J Natl Cancer Inst. 1964, 33:441-56.: 441-456.
34.
Richardson GS, Dickersin GR, Atkins L, MacLaughlin DT, Raam S, Merk LP, Bradley FM: KLE: a cell line with defective estrogen receptor derived from undifferentiated endometrial cancer. Gynecol Oncol. 1984, 17: 213-230. 10.1016/0090-8258(84)90080-5.CrossRefPubMed
35.
Lane EB, Alexander CM: Use of keratin antibodies in tumor diagnosis. Semin Cancer Biol. 1990, 1: 165-179.PubMed
36.
Atkin NB: Modal DNA value and chromosome number in ovarian neoplasia. A clinical and histopathologic assessment. Cancer. 1971, 27: 1064-1073. 10.1002/1097-0142(197105)27:5<1064::AID-CNCR2820270510>3.0.CO;2-K.CrossRefPubMed
37.
Kunzmann R, Holzel F: Karyotype alterations in human ovarian carcinoma cells during long-term cultivation and nude mouse passage. Cancer Genet Cytogenet. 1987, 28: 201-212. 10.1016/0165-4608(87)90206-8.CrossRefPubMed
38.
Gao J, Mazella J, Tang M, Tseng L: Ligand-activated progesterone receptor isoform hPR-A is a stronger transactivator than hPR-B for the expression of IGFBP-1 (insulin-like growth factor binding protein-1) in human endometrial stromal cells. Mol Endocrinol. 2000, 14: 1954-1961. 10.1210/me.14.12.1954.CrossRefPubMed
39.
Arnett-Mansfield RL, deFazio A, Wain GV, Jaworski RC, Byth K, Mote PA, Clarke CL: Relative expression of progesterone receptors A and B in endometrioid cancers of the endometrium. Cancer Res. 2001, 61: 4576-4582.PubMed
40.
Fujimoto J, Ichigo S, Hirose R, Sakaguchi H, Tamaya T: Clinical implication of expression of progesterone receptor form A and B mRNAs in secondary spreading of gynecologic cancers. J Steroid Biochem Mol Biol. 1997, 62: 449-454. 10.1016/S0960-0760(97)00057-5.CrossRefPubMed
41.
Underhill C: CD44: the hyaluronan receptor. J Cell Sci. 1992, 103: 293-298.PubMed
42.
Ferguson SE, Olshen AB, Viale A, Awtrey CS, Barakat RR, Boyd J: Gene expression profiling of tamoxifen-associated uterine cancers: evidence for two molecular classes of endometrial carcinoma. Gynecol Oncol. 2004, 92: 719-725. 10.1016/j.ygyno.2003.10.038.CrossRefPubMed
43.
Graesslin O, Cortez A, Uzan C, Birembaut P, Quereux C, Darai E: Endometrial tumor invasiveness is related to metalloproteinase 2 and tissue inhibitor of metalloproteinase 2 expressions. Int J Gynecol Cancer. 2006, 16: 1911-1917. 10.1111/j.1525-1438.2006.00717.x.CrossRefPubMed
44.
Polette M, Nawrocki-Raby B, Gilles C, Clavel C, Birembaut P: Tumour invasion and matrix metalloproteinases. Crit Rev Oncol Hematol. 2004, 49: 179-186. 10.1016/j.critrevonc.2003.10.008.CrossRefPubMed
45.
Graesslin O, Cortez A, Fauvet R, Lorenzato M, Birembaut P, Darai E: Metalloproteinase-2, -7 and -9 and tissue inhibitor of metalloproteinase-1 and -2 expression in normal, hyperplastic and neoplastic endometrium: a clinical-pathological correlation study. Ann Oncol. 2006, 17: 637-645. 10.1093/annonc/mdj129.CrossRefPubMed
46.
Deligdisch L, Holinka CF: Endometrial carcinoma: two diseases?. Cancer Detect Prev. 1987, 10: 237-246.PubMed
47.
Cho JY, Kim JH, Lee YH, Chung KY, Kim SK, Gong SJ, You NC, Chung HC, Roh JK, Kim BS: Correlation between K-ras gene mutation and prognosis of patients with nonsmall cell lung carcinoma. Cancer. 1997, 79: 462-467. 10.1002/(SICI)1097-0142(19970201)79:3<462::AID-CNCR6>3.0.CO;2-K.CrossRefPubMed
48.
Barbacid M: ras genes. Annu Rev Biochem. 1987, 56:779-827.: 779-827. 10.1146/annurev.bi.56.070187.004023.CrossRef
49.
Bunone G, Briand PA, Miksicek RJ, Picard D: Activation of the unliganded estrogen receptor by EGF involves the MAP kinase pathway and direct phosphorylation. EMBO J. 1996, 15: 2174-2183.PubMedCentralPubMed
50.
Kato S, Endoh H, Masuhiro Y, Kitamoto T, Uchiyama S, Sasaki H, Masushige S, Gotoh Y, Nishida E, Kawashima H, Metzger D, Chambon P: Activation of the estrogen receptor through phosphorylation by mitogen-activated protein kinase. Science. 1995, 270: 1491-1494. 10.1126/science.270.5241.1491.CrossRefPubMed
51.
Niederacher D, An HX, Cho YJ, Hantschmann P, Bender HG, Beckmann MW: Mutations and amplification of oncogenes in endometrial cancer. Oncology. 1999, 56: 59-65. 10.1159/000011931.CrossRefPubMed
52.
53.
Lee JO, Yang H, Georgescu MM, Di Cristofano A, Maehama T, Shi Y, Dixon JE, Pandolfi P, Pavletich NP: Crystal structure of the PTEN tumor suppressor: implications for its phosphoinositide phosphatase activity and membrane association. Cell. 1999, 99: 323-334. 10.1016/S0092-8674(00)81663-3.CrossRefPubMed
54.
Rizo J, Sudhof TC: C2-domains, structure and function of a universal Ca2+-binding domain. J Biol Chem. 1998, 273: 15879-15882. 10.1074/jbc.273.26.15879.CrossRefPubMed
55.
Georgescu MM, Kirsch KH, Akagi T, Shishido T, Hanafusa H: The tumor-suppressor activity of PTEN is regulated by its carboxyl-terminal region. Proc Natl Acad Sci U S A. 1999, 96: 10182-10187. 10.1073/pnas.96.18.10182.PubMedCentralCrossRefPubMed
56.
Davies MA, Lu Y, Sano T, Fang X, Tang P, LaPushin R, Koul D, Bookstein R, Stokoe D, Yung WK, Mills GB, Steck PA: Adenoviral transgene expression of MMAC/PTEN in human glioma cells inhibits Akt activation and induces anoikis. Cancer Res. 1998, 58: 5285-5290.PubMed
57.
Myers MP, Pass I, Batty IH, Van der KJ, Stolarov JP, Hemmings BA, Wigler MH, Downes CP, Tonks NK: The lipid phosphatase activity of PTEN is critical for its tumor supressor function. Proc Natl Acad Sci U S A. 1998, 95: 13513-13518. 10.1073/pnas.95.23.13513.PubMedCentralCrossRefPubMed
58.
Gagnon V, St Germain ME, Parent S, Asselin E: Akt activity in endometrial cancer cells: regulation of cell survival through cIAP-1. Int J Oncol. 2003, 23: 803-810.PubMed
59.
St Germain ME, Gagnon V, Parent S, Asselin E: Regulation of COX-2 protein expression by Akt in endometrial cancer cells is mediated through NF-kappaB/IkappaB pathway. Mol Cancer. 2004, 3:7.: 7-10.1186/1476-4598-3-7.CrossRef
60.
St Germain ME, Gagnon V, Mathieu I, Parent S, Asselin E: Akt regulates COX-2 mRNA and protein expression in mutated-PTEN human endometrial cancer cells. Int J Oncol. 2004, 24: 1311-1324.PubMed
Metadata
Title
Characterization of EN-1078D, a poorly differentiated human endometrial carcinoma cell line: a novel tool to study endometrial invasion in vitro
Authors
Marie-Claude Dery
Celine Van Themsche
Diane Provencher
Anne-Marie Mes-Masson
Eric Asselin
Publication date
01-12-2007
Publisher
BioMed Central
Published in
Reproductive Biology and Endocrinology / Issue 1/2007
Electronic ISSN: 1477-7827
DOI
https://doi.org/10.1186/1477-7827-5-38

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