title: Cell Models for the Investigation of the Role of the Mucin MUC1 Extracellular Domain in Metastasizing.
abstract: The speculations on the role of MUC1, a substance which is overexpressed in glandular cancer cells, on the metastatic potential of such cells are rooted in data that seem to indicate that cell malignization correlates with a change from the apical localization of mucin MUC1 to a peripheral one. Nonetheless, the role of MUC1 in cancer metastasizing remains far from clear. The major hurdle remains the absence of adequate cell models. The aim of the present study was to create cell models that present different fragments of the human mucin MUC1 extracellular domain on their surface. Genetic constructions were generated on the basis of the plasmid vector pEGFP-N3. These constructions contain fusion genes coding for chimeric proteins composed of different combinations of mucin MUC1 functional domains and identification markers (FLAG-epitope, located at the N-terminus, and EGFP, located at the C-terminus of the chimeric proteins). These constructions were used for a stable transformation of HT-29 human cancer cells. The transformants obtained were characterized by flow cytometry. The low expression level of endogenous mucin MUC1 and the high expression level of recombinant proteins were confirmed by real-time PCR. The microscopic examination of the transformed cells confirmed the membrane localization of the fusion proteins. The resulting cell models could be used to investigate the role of the mucin MUC1 domains in cancer cell metastasizing. The obtained cells are used as an applicable model of MUC1-expressing cancers and might be used to study the role of different functional fragments of mucin MUC1 in metastasizing.
authors.affiliation: Department of Molecular Biology, M.V. Lomonosov Moscow State University, Leninskie Gory, 1/12, 119899, Moscow, Russia ; A.N. Bach Institute of Biochemistry, Russian Academy of Science, Leninsky prospekt, 33/2, 119071, Moscow, Russia.
authors.forename: M S
authors.initials: MS
authors.lastname: Syrkina
authors.affiliation: Department of Molecular Biology, M.V. Lomonosov Moscow State University, Leninskie Gory, 1/12, 119899, Moscow, Russia.
authors.forename: M A
authors.initials: MA
authors.lastname: Rubtsov
authors.affiliation: Department of Cell Biology and Histology, M.V. Lomonosov Moscow State University, Leninskie Gory, 1/12, 119899, Moscow, Russia.
authors.forename: D M
authors.initials: DM
authors.lastname: Potashnikova
authors.affiliation: Department of Molecular Biology, M.V. Lomonosov Moscow State University, Leninskie Gory, 1/12, 119899, Moscow, Russia.
authors.forename: Y D
authors.initials: YD
authors.lastname: Kondratenko
authors.affiliation: Bioengineering Department, M.V. Lomonosov Moscow State University, Leninskie Gory, 1/12, 119899, Moscow, Russia.
authors.forename: A A
authors.initials: AA
authors.lastname: Dokrunova
authors.affiliation: A.N. Bach Institute of Biochemistry, Russian Academy of Science, Leninsky prospekt, 33/2, 119071, Moscow, Russia.
authors.forename: V P
authors.initials: VP
authors.lastname: Veiko
keywords: HT-29; MUC1; cancer; cell models; metastasis; mucin
references.citation: J Biol Chem. 1996 Feb 2;271(5):2443-7
references.pmid: 8576205
references.citation: J Biol Chem. 1990 Sep 5;265(25):15286-93
references.pmid: 1697589
references.citation: Hum Pathol. 1995 Apr;26(4):432-9
references.pmid: 7705823
references.citation: Br J Cancer. 2003 Jul 21;89(2):284-90
references.pmid: 12865918
references.citation: Oncogene. 2003 Mar 6;22(9):1324-32
references.pmid: 12618757
references.citation: Clin Exp Metastasis. 2005;22(6):475-83
references.pmid: 16320110
references.citation: J Biol Chem. 1996 Jan 26;271(4):2332-40
references.pmid: 8567697
references.citation: J Cell Biochem. 2009 Nov 1;108(4):802-15
references.pmid: 19711367
references.citation: J Biol Chem. 2004 Jul 9;279(28):29386-90
references.pmid: 15169768
references.citation: Clin Exp Metastasis. 1991 May-Jun;9(3):245-57
references.pmid: 1676353
references.citation: J Biol Chem. 1992 Mar 25;267(9):6171-7
references.pmid: 1556125
references.citation: J Natl Cancer Inst. 1985 Sep;75(3):447-54
references.pmid: 2863413
references.citation: J Pathol. 1997 Nov;183(3):311-7
references.pmid: 9422987
references.citation: Exp Eye Res. 2006 Sep;83(3):493-501
references.pmid: 16631167
references.citation: Mol Biol Cell. 2005 Sep;16(9):4386-97
references.pmid: 15987741
references.citation: Oncogene. 2006 Jan 5;25(1):20-31
references.pmid: 16158055
references.citation: Oncogene. 2010 May 20;29(20):2893-904
references.pmid: 20348949
references.citation: Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7390-7
references.pmid: 7519775
references.citation: Trends Biochem Sci. 1992 Sep;17(9):359-63
references.pmid: 1412714
references.citation: Bioorg Khim. 2000 Jun;26(6):423-32
references.pmid: 10923190
references.citation: J Gastroenterol. 2003;38(12):1162-6
references.pmid: 14714254
references.citation: Oncogene. 2011 Mar 24;30(12):1449-59
references.pmid: 21102519
references.citation: Cancer Cell. 2005 Feb;7(2):167-78
references.pmid: 15710329
references.citation: Mol Cancer. 2011 Jul 28;10:93
references.pmid: 21798038
references.citation: J Biol Chem. 2001 Apr 20;276(16):13057-64
references.pmid: 11278868
references.citation: J Biol Chem. 2001 Sep 21;276(38):35239-42
references.pmid: 11483589
references.citation: Br J Cancer. 1999 Aug;80(12):1867-74
references.pmid: 10471033
references.citation: Breast Cancer Res Treat. 2005 Mar;90(1):77-84
references.pmid: 15770530
references.citation: J Biol Chem. 1997 Sep 26;272(39):24198-202
references.pmid: 9305871
references.citation: Int J Cancer. 1989 Oct 15;44(4):691-6
references.pmid: 2477336
references.citation: Dis Colon Rectum. 1997 Apr;40(4):420-31
references.pmid: 9106690