title: Loss of the BRCA1-interacting helicase BRIP1 results in abnormal mammary acinar morphogenesis.
abstract: BRIP1 is a DNA helicase that directly interacts with the C-terminal BRCT repeat of the breast cancer susceptibility protein BRCA1 and plays an important role in BRCA1-dependent DNA repair and DNA damage-induced checkpoint control. Recent studies implicate BRIP1 as a moderate/low-penetrance breast cancer susceptibility gene. However, the phenotypic effects of BRIP1 dysfunction and its role in breast cancer tumorigenesis remain unclear. To explore the function of BRIP1 in acinar morphogenesis of mammary epithelial cells, we generated BRIP1-knockdown MCF-10A cells by short hairpin RNA (shRNA)-mediated RNA interference and examined its effect in a three-dimensional culture model. Genome-wide gene expression profiling by microarray and quantitative RT-PCR were performed to identify alterations in gene expression in BRIP1-knockdown cells compared with control cells. The microarray data were further investigated using the pathway analysis and Gene Set Enrichment Analysis (GSEA) for pathway identification. BRIP1 knockdown in non-malignant MCF-10A mammary epithelial cells by RNA interference induced neoplastic-like changes such as abnormal cell adhesion, increased cell proliferation, large and irregular-shaped acini, invasive growth, and defective lumen formation. Differentially expressed genes, including MCAM, COL8A1, WIPF1, RICH2, PCSK5, GAS1, SATB1, and ELF3, in BRIP1-knockdown cells compared with control cells were categorized into several functional groups, such as cell adhesion, polarity, growth, signal transduction, and developmental process. Signaling-pathway analyses showed dysregulation of multiple cellular signaling pathways, involving LPA receptor, Myc, Wnt, PI3K, PTEN as well as DNA damage response, in BRIP1-knockdown cells. Loss of BRIP1 thus disrupts normal mammary morphogenesis and causes neoplastic-like changes, possibly via dysregulating multiple cellular signaling pathways functioning in the normal development of mammary glands.
authors.affiliation: Radiobiology for Children's Health Program, Research Center for Radiation Protection, National Institute of Radiological Sciences, Chiba, Japan ; Radiation Effect Accumulation and Prevention Project, Fukushima Project Headquarters, National Institute of Radiological Sciences, Chiba, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Daino
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Imaoka
authors.forename: Takamitsu
authors.initials: T
authors.lastname: Morioka
authors.forename: Shusuke
authors.initials: S
authors.lastname: Tani
authors.forename: Daisuke
authors.initials: D
authors.lastname: Iizuka
authors.forename: Mayumi
authors.initials: M
authors.lastname: Nishimura
authors.forename: Yoshiya
authors.initials: Y
authors.lastname: Shimada
mesh_terms: D061354:Acinar Cells / Q000378:metabolism* / Q000473:pathology*; D000818:Animals; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D016000:Cluster Analysis; D004268:DNA-Binding Proteins / Q000172:deficiency* / Q000235:genetics; D051856:Fanconi Anemia Complementation Group Proteins; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D064593:Heterografts; D006801:Humans; D042361:Mammary Glands, Human / Q000473:pathology*; D051379:Mice; D009024:Morphogenesis / Q000235:genetics*; D020365:RNA Helicases / Q000172:deficiency* / Q000235:genetics; D034622:RNA Interference; D015398:Signal Transduction
chemical_list: D004268:DNA-Binding Proteins; D051856:Fanconi Anemia Complementation Group Proteins; C000618935:BRIP1 protein, human; D020365:RNA Helicases
references.citation: Organogenesis. 2008 Apr;4(2):116-22
references.pmid: 19279723
references.citation: Cold Spring Harb Symp Quant Biol. 2005;70:35-41
references.pmid: 16869736
references.citation: Int J Exp Pathol. 2006 Aug;87(4):289-96
references.pmid: 16875494
references.citation: Nat Genet. 2003 Jul;34(3):267-73
references.pmid: 12808457
references.citation: PLoS One. 2012;7(12):e51786
references.pmid: 23251624
references.citation: J Cell Biol. 2009 Mar 9;184(5):721-36
references.pmid: 19273615
references.citation: Cancer Res. 2009 May 15;69(10):4545-52
references.pmid: 19417140
references.citation: Science. 2003 Oct 24;302(5645):639-42
references.pmid: 14576433
references.citation: J Cell Biol. 2004 Feb 16;164(4):603-12
references.pmid: 14769856
references.citation: Oncogene. 1997 Apr 3;14(13):1617-22
references.pmid: 9129154
references.citation: Nat Genet. 1999 May;22(1):37-43
references.pmid: 10319859
references.citation: Am J Pathol. 1997 Sep;151(3):745-51
references.pmid: 9284823
references.citation: Nat Genet. 2006 Nov;38(11):1239-41
references.pmid: 17033622
references.citation: Future Oncol. 2011 Feb;7(2):253-61
references.pmid: 21345144
references.citation: Biochem Soc Symp. 1998;63:211-21
references.pmid: 9513725
references.citation: Endocrinology. 2011 Dec;152(12):5041-52
references.pmid: 21971156
references.citation: Genome Biol. 2009;10(6):226
references.pmid: 19589182
references.citation: Mol Pharmacol. 2000 Dec;58(6):1188-96
references.pmid: 11093753
references.citation: Cancer Res. 2005 Dec 1;65(23):10992-1000
references.pmid: 16322248
references.citation: Carcinogenesis. 2010 Sep;31(9):1694-701
references.pmid: 20616149
references.citation: Oncogene. 2006 Apr 6;25(15):2245-53
references.pmid: 16462773
references.citation: J Natl Cancer Inst. 1999 Jun 2;91(11):943-9
references.pmid: 10359546
references.citation: Proc Natl Acad Sci U S A. 2004 Feb 24;101(8):2357-62
references.pmid: 14983014
references.citation: Expert Rev Anticancer Ther. 2008 Oct;8(10):1689-98
references.pmid: 18925859
references.citation: Methods. 2003 Jul;30(3):256-68
references.pmid: 12798140
references.citation: Nature. 2008 Mar 13;452(7184):187-93
references.pmid: 18337816
references.citation: Nature. 2005 Apr 14;434(7035):864-70
references.pmid: 15829956
references.citation: Oncogene. 2007 Dec 10;26(56):7799-808
references.pmid: 18066093
references.citation: Cancer Cell Int. 2009 Jul 30;9:18
references.pmid: 19642980
references.citation: Breast Cancer Res. 1999;1(1):14-7
references.pmid: 11250676
references.citation: Bioessays. 2004 Aug;26(8):870-81
references.pmid: 15273989
references.citation: Clin Cancer Res. 2008 Jul 15;14(14):4672-80
references.pmid: 18628483
references.citation: Nat Rev Cancer. 2006 Mar;6(3):184-92
references.pmid: 16453012
references.citation: Proc Natl Acad Sci U S A. 2005 Jun 28;102(26):9176-81
references.pmid: 15967981
references.citation: Genes Cancer. 2010 Jun;1(6):629-40
references.pmid: 21779462
references.citation: Cell. 2001 Apr 6;105(1):149-60
references.pmid: 11301010
references.citation: Am J Hum Genet. 1997 Mar;60(3):486-95
references.pmid: 9042907
references.citation: Proc Natl Acad Sci U S A. 2005 Oct 25;102(43):15545-50
references.pmid: 16199517
references.citation: Am J Hum Genet. 1997 May;60(5):1031-40
references.pmid: 9150150
references.citation: Science. 2008 Mar 7;319(5868):1352-5
references.pmid: 18323444
references.citation: FASEB J. 1998 Nov;12(14):1541-50
references.pmid: 9806763
references.citation: Cancer Cell. 2009 Jun 2;15(6):539-50
references.pmid: 19477432
references.citation: Breast Cancer Res. 2008;10(3):208
references.pmid: 18557994
references.citation: Cancer Res. 1998 Mar 1;58(5):1004-12
references.pmid: 9500463
references.citation: Nat Rev Cancer. 2011 Dec 23;12(1):68-78
references.pmid: 22193408
references.citation: Breast Cancer Res. 2004;6(6):R640-6
references.pmid: 15535846
references.citation: Dev Cell. 2011 Dec 13;21(6):1077-91
references.pmid: 22137763
references.citation: Am J Hum Genet. 1997 May;60(5):1013-20
references.pmid: 9150148
references.citation: Am J Hum Genet. 2003 May;72(5):1117-30
references.pmid: 12677558
references.citation: Nature. 2005 Apr 14;434(7035):907-13
references.pmid: 15829965
references.citation: Hum Mol Genet. 2003 Apr 1;12 Spec No 1:R113-23
references.pmid: 12668603