Repression of p63 and induction of EMT by mutant Ras in mammary epithelial cells.
The p53-related transcription factor p63 is required for maintenance of epithelial cell differentiation. We found that activated forms of the Harvey Rat Sarcoma Virus GTPase (H-RAS) and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) oncogenes strongly repress expression of ∆Np63α, the predominant p63 isoform in basal mammary epithelial cells. This regulation occurs at the transcriptional level, and a short region of the ∆Np63 promoter is sufficient for repression induced by H-RasV12. The suppression of ∆Np63α expression by these oncogenes concomitantly leads to an epithelial-to-mesenchymal transition (EMT). In addition, the depletion of ∆Np63α alone is sufficient to induce EMT. Both H-RasV12 expression and ∆Np63α depletion induce individual cell invasion in a 3D collagen gel in vitro system, thereby demonstrating how Ras can drive the mammary epithelial cell state toward greater invasive ability. Together, these results suggest a pathway by which RAS and PIK3CA oncogenes induce EMT through regulation of ∆Np63α.
Department of Biological Sciences, Columbia University, New York, NY 10027.
Kathryn E
KE
Yoh
Department of Biological Sciences, Columbia University, New York, NY 10027.
Kausik
K
Regunath
Department of Chemistry, Columbia University, New York, NY 10027.
Asja
A
Guzman
Department of Food and Nutritional Sciences, College of Human Ecology, Yonsei University, Seoul 03722, South Korea.
Seung-Min
SM
Lee
Department of Biological Sciences, Columbia University, New York, NY 10027.
Neil T
NT
Pfister
Department of Biological Sciences, Columbia University, New York, NY 10027.
Olutosin
O
Akanni
Department of Chemistry, Columbia University, New York, NY 10027.
Laura J
LJ
Kaufman
Department of Biological Sciences, Columbia University, New York, NY 10027; clp3@columbia.edu mrp6@columbia.edu.
Carol
C
Prives
Department of Biological Sciences, Columbia University, New York, NY 10027; clp3@columbia.edu mrp6@columbia.edu.
Ron
R
Prywes
D001483:Base Sequence; D001665:Binding Sites; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism*; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D008954:Models, Biological; D009154:Mutation; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D017384:Sequence Deletion; D015398:Signal Transduction; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018631:ras Proteins / Q000235:genetics*
D034741:RNA, Small Interfering; C115051:TP63 protein, human; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D018631:ras Proteins
H-Ras; breast cancer; epithelial mesenchymal transition; p63; transcriptional repression
Nat Rev Cancer. 2013 Feb;13(2):97-110
23344542
Bioinformatics. 2015 Jan 15;31(2):166-9
25260700
Nature. 2015 Nov 26;527(7579):472-6
26560033
Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3255-60
17360634
Cancer Res. 2012 May 15;72(10):2457-67
22589270
Nat Cell Biol. 2014 Oct;16(10):942-50, 1-7
25173976
J Biol Chem. 2011 Feb 4;286(5):3915-24
21127042
Acta Naturae. 2015 Apr-Jun;7(2):17-28
26085941
Expert Rev Anticancer Ther. 2011 Feb;11(2):263-75
21342044
Proc Natl Acad Sci U S A. 1981 Jun;78(6):3328-32
6267583
Mol Cell Biol. 2009 Jul;29(14 ):3953-63
19451233
Nat Rev Mol Cell Biol. 2003 Aug;4(8):657-65
12923528
Nat Methods. 2014 Feb;11(2):119-20
24645192
BMC Bioinformatics. 2011 Aug 04;12:323
21816040
Nature. 1999 Apr 22;398(6729):708-13
10227293
Nat Methods. 2014 Feb;11(2):117
24645191
Oncogene. 2000 Mar 9;19(11):1379-85
10723128
Tissue Eng Part C Methods. 2008 Sep;14(3):261-71
18694322
J Clin Oncol. 2009 Mar 10;27(8):1160-7
19204204
Cell. 2009 Dec 24;139(7):1327-41
20064378
Nucleic Acids Res. 2005 Nov 24;33(20):6566-78
16314317
J Cell Biol. 2010 Jan 11;188(1):11-9
19951899
Genes Dev. 2004 Jan 15;18(2):126-31
14729569
Cell. 2009 Nov 25;139(5):871-90
19945376
PLoS One. 2011;6(12 ):e28688
22205962
J Clin Invest. 2009 Jun;119(6):1420-8
19487818
Proc Natl Acad Sci U S A. 2012 Sep 18;109(38):15312-7
22949650
Cancer Res. 2007 Jan 15;67(2):501-10
17234757
Genome Biol. 2010;11(10):R106
20979621
J Invest Dermatol. 2006 Jul;126(7):1469-79
16645595
Science. 1987 Jan 9;235(4785):177-82
3798106
Exp Cell Res. 2006 Apr 1;312(6):695-706
16406339
Bioinformatics. 2009 May 1;25(9):1105-11
19289445
Nat Methods. 2009 Apr;6(4):239-40
19333237
J Pathol. 2002 Dec;198(4):417-27
12434410
Nucleic Acids Res. 1990 Jun 25;18(12):3587-96
2194165
J Biol Chem. 2008 May 30;283(22):14910-4
18411277
Nucleic Acids Res. 2009 Jan;37(1):1-13
19033363
Nucleic Acids Res. 2011 Jan;39(Database issue):D945-50
20952405
Bioinformatics. 2014 Aug 1;30(15):2114-20
24695404
Cancer Res. 2009 Mar 15;69(6):2191-4
19276389
Cancer Res. 1997 May 15;57(10):2061-70
9158006
Am J Pathol. 2002 Oct;161(4):1199-206
12368193
Nature. 1999 Apr 22;398(6729):714-8
10227294
Cell. 1991 Oct 18;67(2):403-10
1913826
Bioinformatics. 2009 Aug 15;25(16):2078-9
19505943
Nat Biotechnol. 2011 Jan;29(1):79-83
21131983
Cell. 2011 Nov 23;147(5):992-1009
22118458
Mol Cell. 1998 Sep;2(3):305-16
9774969
Oncogene. 2010 Aug 19;29(33):4715-24
20562907
BMC Med Genomics. 2012 Oct 04;5:44
23035882
Nature. 2015 Nov 26;527(7579):525-30
26560028
PLoS One. 2010 Sep 03;5(9):e12336
20838429
Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1624-8
7878029
Proc Natl Acad Sci U S A. 2000 May 9;97(10 ):5462-7
10805802
Stat Med. 1990 Jul;9(7):811-8
2218183
Curr Top Dev Biol. 2014;107:133-60
24439805
Sci Signal. 2013 Apr 02;6(269):pl1
23550210
Cell Cycle. 2006 Sep;5(17 ):1996-2004
16931914
Oncogene. 2014 Apr 3;33(14 ):1755-63
23604123
Nat Cell Biol. 2006 Jun;8(6):551-61
16715076
Cancer Res. 1990 Sep 15;50(18):6075-86
1975513
Mol Cell Biol. 2013 Aug;33(15):2930-49
23716601
Cancer Biol Ther. 2004 Aug;3(8):772-5
15254419
Science. 2004 Apr 23;304(5670):554
15016963
J Biol Chem. 2013 Feb 1;288(5):3275-88
23239884
Virchows Arch. 2015 Jul;467(1):67-70
25850754
Oncogene. 1987;1(4):423-30
3330785
Nucleic Acids Res. 2015 Jan;43(Database issue):D812-7
25392408
Cell Stem Cell. 2010 Sep 3;7(3):403-17
20804975
Nature. 1998 Mar 12;392(6672):190-3
9515965
Cell. 1987 Sep 11;50(6):863-72
3476205
Proc Natl Acad Sci U S A. 2005 Dec 20;102(51):18443-8
16339315
Development. 2012 Feb;139(4):772-82
22274697
Mod Pathol. 1995 Jun;8(5):515-20
7675770
Nature. 2012 Oct 4;490(7418):61-70
23000897
J Cell Biol. 2002 Jan 21;156(2):299-313
11790801
J Biol Chem. 2002 Oct 18;277(42):39209-16
12161443
Cell. 1983 Nov;35(1):79-87
6313230
Nat Cell Biol. 2008 May;10 (5):593-601
18376396
Genes Dev. 1996 Oct 1;10(19):2462-77
8843198
Int J Cancer. 2002 Dec 20;102(6):580-3
12447998
Am J Surg Pathol. 2001 Aug;25(8):1054-60
11474290
J Clin Invest. 2007 May;117(5):1370-80
17446929
Proc Natl Acad Sci U S A. 1985 Nov;82(21):7409-13
2414776
Cell. 2009 Apr 3;137(1):87-98
19345189
Mol Cell. 2001 Jun;7(6):1267-78
11430829
Histol Histopathol. 2004 Apr;19(2):465-71
15024707
Nat Protoc. 2009;4(1):44-57
19131956
J Natl Cancer Inst. 1993 Nov 3;85(21):1725-32
8411256
Nucleic Acids Res. 2012 Jan;40(2):499-510
21917857
Nat Med. 2009 Aug;15(8):907-13
19648928
Sci Signal. 2011 Sep 13;4(191):tr7
21934110
Breast Cancer Res Treat. 2010 Aug;122(3):765-75
19898932
Front Endocrinol (Lausanne). 2015 Apr 28;6:51
25972840
BMC Genomics. 2008 Oct 16;9:488
18925949
Proc Natl Acad Sci U S A. 2005 Jan 18;102(3):802-7
15647370
Clin Cancer Res. 2008 Mar 1;14(5):1368-76
18316557
Int J Cancer. 2008 Jul 1;123(1):236-40
18398844
Virchows Arch. 2003 Aug;443(2):122-32
12884041
J Biomol Screen. 2006 Dec;11(8):922-32
16973921
Nat Med. 2013 Nov;19(11):1450-64
24202397