Establishment and characterization of cell lines derived from uterine malignant mixed Müllerian tumor.

OBJECTIVE We report the establishment and characterization of three new cell lines derived from uterine malignant mixed müllerian tumor (MMMT). METHODS Three uterine MMMT cell lines from primary tumors of Korean patients were cultured and the involved cell morphology, growth properties, DNA profiles, immunohistochemical properties, tumor-associated antigen secretion, and genetic alterations of related oncogenes and tumor suppressor genes were studied as well. RESULTS Three MMMT cell lines were successfully established including one homologous tumor SNU-539 and two heterologous tumors SNU-685 and SNU-1077. All lines showed substrate adherence and high viability and were proven by DNA fingerprinting analysis to be unique. Contamination by mycoplasma and bacteria was excluded. SNU-539 and SNU-1077 cells stained positively for both epithelial and mesenchymal antigens, while SNU-685 cells only stained positively for mesenchymal antigens. The level of secretion of tumor-associated antigens, CA125 and CEA, was shown to be undetectable in all three lines. One missense mutation from AAC to GAC at codon 239 of exon 7 in the p53 gene was identified in SNU-539. CONCLUSIONS These newly established and characterized permanent uterine MMMT cell lines might be regarded as valuable resources for a multitude of in vitro investigations, which should be used for clarifying the obscure histogenetic origin and understanding the biological behavior of this aggressive tumor.

[1]  R. Bast,et al.  The p53 tumor suppressor gene frequently is altered in gynecologic cancers. , 1994 .

[2]  W B OBER,et al.  UTERINE SARCOMAS: HISTOGENESIS AND TAXONOMY , 1959 .

[3]  Y. Kaneko,et al.  Two cell lines established from mixed Müllerian tumors of the uterus morphologic, immunocytochemical, and cytogenetic analyses , 1992, Cancer.

[4]  R. Ian Freshney,et al.  Culture of Animal Cells , 1983 .

[5]  H. Iwasaki,et al.  Characteristics of cloned cells of mixed müllerian tumor of the human uterus carcinoma cells showing myogenic differentiation in vitro , 1993, Cancer.

[6]  M. Williamson,et al.  p16 (CDKN2) is a major deletion target at 9p21 in bladder cancer. , 1995, Human molecular genetics.

[7]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[8]  K. Geisinger,et al.  Malignant mixed müllerian tumors an ultrastructural and immunohistochemical analysis with histogenetic considerations , 1987, Cancer.

[9]  B. Scheithauer,et al.  Clinicopathologic analysis of uterine malignant mixed müllerian tumors. , 1989, Gynecologic oncology.

[10]  T. Sekiya,et al.  Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. , 1989, Genomics.

[11]  A. Gown,et al.  The expression of epidermal growth factor receptor, HER-2/Neu, p53, and Ki-67 antigen in uterine malignant mixed mesodermal tumors and adenosarcoma. , 1996, Gynecologic oncology.

[12]  E. O. Williamson,et al.  Malignant mixed müllerian tumors of the uterus , 1972, Cancer.

[13]  Y. Shimosato,et al.  EXPRESSION OF VARIOUS ANTIGENS BY DIFFERENT COMPONENTS OF UTERINE MIXED MÜLLERIAN TUMORS: An Immunohistochemical Study , 1988, Acta pathologica japonica.

[14]  A. Gazdar,et al.  CHARACTERISTICS OF COLORECTAL CARCINOMA CELL LINES paraffin embedded sections of xenografts , 2006 .

[15]  I. Ishiwata,et al.  Establishment and characterization of two human mixed mesodermal tumor cell lines from the same patient. , 1984, Journal of the National Cancer Institute.

[16]  Y. Nakamura,et al.  Germline mutations of hMLH1 and hMSH2 genes in Korean hereditary nonpolyposis colorectal cancer. , 1996, Journal of the National Cancer Institute.

[17]  T. Sekiya,et al.  Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[18]  B. Sikic,et al.  Development and characterization of a human sarcoma cell line, MES-SA, sensitive to multiple drugs. , 1983, Cancer research.

[19]  F. Collins,et al.  Mutations in the p53 gene occur in diverse human tumour types , 1989, Nature.

[20]  J. Boyd,et al.  Mutation and over‐expression of the P53 tumor suppressor gene frequently occurs in uterine and ovarian sarcomas , 1994, Obstetrics and gynecology.

[21]  G. Fletcher,et al.  Malignant mixed müllerian tumors of the uterus , 1984, Cancer.

[22]  H. Taylor,et al.  Mesenchymal tumors of the uterus: III. A clinical and pathologic study of 31 carcinosarcomas , 1966 .

[23]  C. Harris,et al.  Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis. , 1994, Cancer research.

[24]  Y. Nakamura,et al.  Genetic instability in pancreatic cancer and poorly differentiated type of gastric cancer. , 1993, Cancer research.

[25]  E. M. Gilder,et al.  Handbuch der speziellen pathologischen Anatomie und Histologie , 1972 .

[26]  M. Feldstein,et al.  Uterine sarcomas. Natural history, treatment and prognosis , 1978, Cancer.

[27]  J. Minna,et al.  Growth of cell lines and clinical specimens of human non-small cell lung cancer in a serum-free defined medium. , 1986, Cancer research.

[28]  H. Taylor,et al.  Mesenchymal tumors of the uterus. II. A clinical and pathologic study of 31 mixed mesodermal tumors. , 1966, Obstetrics and gynecology.

[29]  Graham Jb,et al.  Carcinosarcoma and mixed mesodermal tumor of the uterine corpus. Review of 49 cases. , 1970 .

[30]  K. Shroyer,et al.  Alteration of the p53 tumor suppressor gene and activation of c-K-ras-2 protooncogene in endometrial adenocarcinoma from Colorado. , 1995, American journal of clinical pathology.

[31]  M. Kelley,et al.  Mutation of p53 gene in hepatocellular carcinoma cell lines with HBX DNA , 1996, International Journal of Cancer.

[32]  K. Klaamas,et al.  IgG immune response to Helicobacter pylori antigens in patients with gastric cancer as defined by ELISA and immunoblotting , 1996, International journal of cancer.

[33]  H. Ishikawa,et al.  Histogenesis and culture of human uterine carcinosarcoma. , 1981, Cancer research.

[34]  Steven E. Bayer,et al.  A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1. , 1994, Science.

[35]  R. White,et al.  Microsatellite instability in colorectal adenocarcinoma cell lines that have full-length adenomatous polyposis coli protein. , 1995, Cancer research.

[36]  R. Kempson,et al.  Uterine sarcomas: Classification, diagnosis, and prognosis , 1970 .

[37]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[38]  J. Goding,et al.  Monoclonal Antibodies: Principles and Practice , 1996 .

[39]  H. Minaguchi,et al.  Establishment and characterization of carcinosarcoma cell line of the human uterus , 1993, Cancer.